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do Nascimento RG, da Conceição MPF, de Bastos DR, de Toledo Osorio CAB, López RVM, Reis EM, Cerqueira OLD. Prognostic value of Maspin protein level in patients with triple negative breast cancer. Sci Rep 2024; 14:15982. [PMID: 38987610 PMCID: PMC11237076 DOI: 10.1038/s41598-024-53870-y] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/13/2022] [Accepted: 02/06/2024] [Indexed: 07/12/2024] Open
Abstract
The search for prognostic markers in breast cancer has bumped into a typical feature of these tumors, intra and intertumoral heterogeneity. Changes in the expression profile, localization of these proteins or shedding to the surrounding stroma can be useful in the search for new markers. In this context, classification by molecular subtypes can bring perspectives for both diagnosis and screening for appropriate treatments. However, the Triple Negative (TN) subtype, which is already the one with the worst prognosis, lacks appropriate and consistent molecular markers. In this work, we analyzed 346 human breast cancer samples in tissue microarrays (TMA) from cases diagnosed with invasive breast carcinoma to assess the expression and localization pattern of Maspin and their correlation with clinical parameters. To complement our findings, we also used TCGA data to analyze the mRNA levels of these respective genes. Our data suggests that the TN subtype demonstrates a higher level of cytoplasmic Maspin compared to the other subtypes. Maspin transcript levels follow the same trend. However, TN patients with lower Maspin expression tend to have worse overall survival and free-survival metastasis rates. Finally, we used Maspin expression data to verify possible relationships with the clinicopathological information of our cohort. Our univariate analyses indicate that Maspin is related to the expression of estrogen receptor (ER) and progesterone receptor (PR). Furthermore, Maspin expression levels also showed correlation with Scarff-Bloom-Richardson (SBR) parameter, and stromal Maspin showed a relationship with lymph node involvement. Our data is not consistently robust enough to categorize Maspin as a prognostic marker. However, it does indicate a change in the expression profile within the TN subtype.
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Affiliation(s)
- Renan Gomes do Nascimento
- Center for Translational Research in Oncology, Cancer Institute of the State of São Paulo (ICESP), Clinical Hospital Faculty of Medicine, University of São Paulo (HCFMUSP), São Paulo, SP, 01246-000, Brazil
- Department of Clinical Pharmacy and Oncology, Hospital São Camilo (HSC), São Paulo, SP, 02401-300, Brazil
| | - Mércia Patrícia Ferreira da Conceição
- Center for Translational Research in Oncology, Cancer Institute of the State of São Paulo (ICESP), Clinical Hospital Faculty of Medicine, University of São Paulo (HCFMUSP), São Paulo, SP, 01246-000, Brazil
| | - Daniel Rodrigues de Bastos
- Center for Translational Research in Oncology, Cancer Institute of the State of São Paulo (ICESP), Clinical Hospital Faculty of Medicine, University of São Paulo (HCFMUSP), São Paulo, SP, 01246-000, Brazil
| | | | - Rossana Verónica Mendoza López
- Center for Translational Research in Oncology, Cancer Institute of the State of São Paulo (ICESP), Clinical Hospital Faculty of Medicine, University of São Paulo (HCFMUSP), São Paulo, SP, 01246-000, Brazil
| | - Eduardo Moraes Reis
- Departmento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP, 05508-900, Brazil
| | - Otto Luiz Dutra Cerqueira
- Center for Translational Research in Oncology, Cancer Institute of the State of São Paulo (ICESP), Clinical Hospital Faculty of Medicine, University of São Paulo (HCFMUSP), São Paulo, SP, 01246-000, Brazil.
- Departmento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP, 05508-900, Brazil.
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Rathod M, Franz H, Beyersdorfer V, Wanuske MT, Leal-Fischer K, Hanns P, Stüdle C, Zimmermann A, Buczak K, Schinner C, Spindler V. DPM1 modulates desmosomal adhesion and epidermal differentiation through SERPINB5. J Cell Biol 2024; 223:e202305006. [PMID: 38477878 PMCID: PMC10937187 DOI: 10.1083/jcb.202305006] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/02/2023] [Revised: 11/30/2023] [Accepted: 01/23/2024] [Indexed: 03/14/2024] Open
Abstract
Glycosylation is essential to facilitate cell-cell adhesion and differentiation. We determined the role of the dolichol phosphate mannosyltransferase (DPM) complex, a central regulator for glycosylation, for desmosomal adhesive function and epidermal differentiation. Deletion of the key molecule of the DPM complex, DPM1, in human keratinocytes resulted in weakened cell-cell adhesion, impaired localization of the desmosomal components desmoplakin and desmoglein-2, and led to cytoskeletal organization defects in human keratinocytes. In a 3D organotypic human epidermis model, loss of DPM1 caused impaired differentiation with abnormally increased cornification, reduced thickness of non-corneal layers, and formation of intercellular gaps in the epidermis. Using proteomic approaches, SERPINB5 was identified as a DPM1-dependent interaction partner of desmoplakin. Mechanistically, SERPINB5 reduced desmoplakin phosphorylation at serine 176, which was required for strong intercellular adhesion. These results uncover a novel role of the DPM complex in connecting desmosomal adhesion with epidermal differentiation.
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Affiliation(s)
- Maitreyi Rathod
- Department of Biomedicine, University of Basel, Basel, Switzerland
- Institute of Anatomy and Experimental Morphology, University Clinic Hamburg-Eppendorf, Hamburg, Germany
| | - Henriette Franz
- Department of Biomedicine, University of Basel, Basel, Switzerland
| | - Vivien Beyersdorfer
- Department of Biomedicine, University of Basel, Basel, Switzerland
- Institute of Anatomy and Experimental Morphology, University Clinic Hamburg-Eppendorf, Hamburg, Germany
| | | | | | - Pauline Hanns
- Department of Biomedicine, University of Basel, Basel, Switzerland
| | - Chiara Stüdle
- Department of Biomedicine, University of Basel, Basel, Switzerland
| | - Aude Zimmermann
- Department of Biomedicine, University of Basel, Basel, Switzerland
| | - Katarzyna Buczak
- Proteomics Core Facility, Biocentre, University of Basel, Basel, Switzerland
| | - Camilla Schinner
- Department of Biomedicine, University of Basel, Basel, Switzerland
- Institute of Functional and Applied Anatomy, Hannover Medical School, Hannover, Germany
| | - Volker Spindler
- Department of Biomedicine, University of Basel, Basel, Switzerland
- Institute of Anatomy and Experimental Morphology, University Clinic Hamburg-Eppendorf, Hamburg, Germany
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Gomarasca M, Lombardi G, Maroni P. SUMOylation and NEDDylation in Primary and Metastatic Cancers to Bone. Front Cell Dev Biol 2022; 10:889002. [PMID: 35465332 PMCID: PMC9020829 DOI: 10.3389/fcell.2022.889002] [Citation(s) in RCA: 13] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/03/2022] [Accepted: 03/25/2022] [Indexed: 12/22/2022] Open
Abstract
Post-translational modifications comprise series of enzymatically-driven chemical modifications, virtually involving the entire cell proteome, that affect the fate of a target protein and, in turn, cell activity. Different classes of modifications can be established ranging from phosphorylation, glycosylation, ubiquitination, acetylation, methylation, lipidation and their inverse reactions. Among these, SUMOylation and NEDDylation are ubiquitin-like multi-enzymatic processes that determine the bound of SUMOs and NEDD8 labels, respectively, on defined amino acidic residues of a specific protein and regulate protein function. As fate-determinants of several effectors and mediators, SUMOylation and NEDDylation play relevant roles in many aspects of tumor cell biology. Bone represents a preferential site of metastasis for solid tumors (e.g., breast and prostate cancers) and the primary site of primitive tumors (e.g., osteosarcoma, chondrosarcoma). Deregulation of SUMOylation and NEDDylation affects different aspects of neoplastic transformation and evolution such as epithelial-mesenchymal transition, adaptation to hypoxia, expression and action of tumor suppressors and oncogenic mediators, and drug resistance. Thereby, they represent potential therapeutic targets. This narrative review aims at describing the involvement and regulation of SUMOylation and NEDDylation in tumor biology, with a specific focus on primary and secondary bone tumors, and to summarize and highlight their potentiality in diagnostics and therapeutic strategies.
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Affiliation(s)
- Marta Gomarasca
- Laboratory of Experimental Biochemistry and Molecular Biology, IRCCS Istituto Ortopedico Galeazzi, Milano, Italy
| | - Giovanni Lombardi
- Laboratory of Experimental Biochemistry and Molecular Biology, IRCCS Istituto Ortopedico Galeazzi, Milano, Italy
- Department of Athletics, Strength and Conditioning, Poznań University of Physical Education, Poznań, Polska
- *Correspondence: Giovanni Lombardi,
| | - Paola Maroni
- Laboratory of Experimental Biochemistry and Molecular Biology, IRCCS Istituto Ortopedico Galeazzi, Milano, Italy
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Sinha KK, Vinay J, Parida S, Singh SP, Dixit M. Association and functional significance of genetic variants present in regulatory elements of SERPINB5 gene in gallbladder cancer. Gene 2022; 808:145989. [PMID: 34624458 DOI: 10.1016/j.gene.2021.145989] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/08/2021] [Accepted: 10/01/2021] [Indexed: 11/04/2022]
Abstract
SERPINB5 is a mammary serine protease inhibitor, which is involved in various cellular functions. The aberrant expression of SERPINB5 is reported in many cancers along with GBC but limited information is available about its role in genetic predisposition for GBC. We carried out case-control study in 206 cases and 219 controls. Promoter SNPs were genotyped by Sanger's sequencing. In-silico promoter analysis and luciferase reporter assay were done to elucidate the role of promoter variants in regulation of SERPINB5 expression. Out of four SNPs, three SERPINB5 promoter variants showed association with GBC in different models. The 'C' allele of variant rs17071138 was found to be significantly associated with GBC (p = 0.017). The 'T' allele of rs3744940 significantly increased the risk for GBC in dominant (p = 0.035) and additive models (p = 0.005). Also, rs3744941 'T' allele increased the risk for GBC by dominant (p = 0.042) as well as additive models (p = 0.016). In-silico promoter analysis and luciferase reporter assay revealed the probable regulatory role of the SERPINB5 promoter variant rs17071138 on the expression. Overall, our study reveals the genetic association of SERPINB5 promoter variants with GBC and possible role of rs17071138 in the regulation of expression.
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Affiliation(s)
- Kirti Kumari Sinha
- National Institute of Science Education and Research, School of Biological Sciences, Bhubaneswar, Odisha 752050, India; Homi Bhabha National Institute, Training School Complex, Anushakti Nagar, Mumbai 400094, India
| | - J Vinay
- National Institute of Science Education and Research, School of Biological Sciences, Bhubaneswar, Odisha 752050, India; Homi Bhabha National Institute, Training School Complex, Anushakti Nagar, Mumbai 400094, India
| | - Suryakant Parida
- Sriram Chandra Bhanja Medical College & Hospital, Department of Gastroenterology, Cuttack, Odisha 753007, India
| | - Shivaram Prasad Singh
- Sriram Chandra Bhanja Medical College & Hospital, Department of Gastroenterology, Cuttack, Odisha 753007, India
| | - Manjusha Dixit
- National Institute of Science Education and Research, School of Biological Sciences, Bhubaneswar, Odisha 752050, India; Homi Bhabha National Institute, Training School Complex, Anushakti Nagar, Mumbai 400094, India.
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Khorsandi L, Farasat M. Zinc oxide nanoparticles enhance expression of maspin in human breast cancer cells. ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH INTERNATIONAL 2020; 27:38300-38310. [PMID: 32621200 DOI: 10.1007/s11356-020-09986-5] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/20/2020] [Accepted: 07/01/2020] [Indexed: 06/11/2023]
Abstract
Toxic and apoptotic impacts of zinc oxide nanoparticle (ZNP) on different cancer cells have been reported. Maspin (a mammary serine protease inhibitor) as a tumor suppressor gene can inhibit tumor growth and metastasis. The expression of maspin is modulated by p53, Bcl-2 family genes, and estrogen receptor α (ER-α). This study aimed to assess the ZNP effects on maspin expression in MCF-7 cells (a breast cancer cell). Experimental groups (ZNP5, ZNP10, and ZNP20) received 5, 10, and 20 μM/mL ZNP for 48 h, respectively. 17-β-estradiol (E2) was used to evaluate the role of ER-α in the anticancer impact of ZNP. Cell viability, Annexin V, migration assay, gene expression, and western blotting methods were applied to evaluate ZNP effects on the MCF-7 cells. ZNP at the concentrations of 10 and 20 μM/mL could significantly decrease the viability and migration rate, and significantly increase apoptosis percentage in the MCF-7 cells. ZNP significantly enhanced mRNA expression and protein level of maspin in MCF-7 cells in a concentration-dependent way. ZNP concentration-dependently elevated mRNA expression and protein level of p53 and Bax while reduced the expression of Bcl-2 and ER-α. E2 promoted cancer cell growth by enhancing survival and migration rates. E2 treatment reduced mRNA expression and protein level of maspin and p53, and elevated Bcl-2 expression. ZNP considerably changed these events induced by E2 in the MCF-7 cells. It is concluded that the maspin overexpression is one of the toxic mechanisms of the ZNP on the ER-α-positive breast cancer cells, and can suppress the migration of these cells.
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Affiliation(s)
- Layasadat Khorsandi
- Cellular and Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
- Department of Anatomical Sciences, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
| | - Maryam Farasat
- Department of Anatomical Sciences, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
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Kretzmann JA, Evans CW, Moses C, Sorolla A, Kretzmann AL, Wang E, Ho D, Hackett MJ, Dessauvagie BF, Smith NM, Redfern AD, Waryah C, Norret M, Iyer KS, Blancafort P. Tumour suppression by targeted intravenous non-viral CRISPRa using dendritic polymers. Chem Sci 2019; 10:7718-7727. [PMID: 31588320 PMCID: PMC6761875 DOI: 10.1039/c9sc01432b] [Citation(s) in RCA: 37] [Impact Index Per Article: 6.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/28/2019] [Accepted: 06/26/2019] [Indexed: 12/22/2022] Open
Abstract
This article demonstrates a fully synthetic strategy enabling CRISPR-mediated activation of tumour suppressor genes in vivo to reduce tumour burden.
Aberrant gene expression is a hallmark of cancer. Although transcription is traditionally considered ‘undruggable’, the development of CRISPR-associated protein 9 (Cas9) systems offers enormous potential to rectify cancer-associated transcriptional abnormalities in malignant cells. However delivery of this technology presents a critical challenge to overcome in order to realize clinical translation for cancer therapy. In this article we demonstrate for the first time, a fully synthetic strategy to enable CRISPR-mediated activation (CRISPRa) of tumour suppressor genes in vivo using a targeted intravenous approach. We show this via highly efficient transcriptional activation of two model tumour suppressor genes, Mammary Serine Protease Inhibitor (MASPIN, SERPINB5) and cysteine-rich 61/connective tissue growth factor/nephroblastoma-overexpressed 6 (CCN6, WISP3), in a mouse model of breast cancer. In particular, we demonstrate that targeted intravenous delivery of can be achieved using a novel nanoscale dendritic macromolecular delivery agent, with negligible toxicity and long lasting therapeutic effects, outlining a targeted effective formulation with potential to treat aggressive malignancies.
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Affiliation(s)
- Jessica A Kretzmann
- School of Molecular Sciences , The University of Western Australia , 35 Stirling Hwy , Crawley , WA 6009 , Australia . .,Harry Perkins Institute of Medical Research , 6 Verdun St , Nedlands , WA 6009 , Australia .
| | - Cameron W Evans
- School of Molecular Sciences , The University of Western Australia , 35 Stirling Hwy , Crawley , WA 6009 , Australia .
| | - Colette Moses
- Harry Perkins Institute of Medical Research , 6 Verdun St , Nedlands , WA 6009 , Australia . .,School of Human Sciences , The University of Western Australia , 35 Stirling Hwy , Crawley , WA 6009 , Australia
| | - Anabel Sorolla
- Harry Perkins Institute of Medical Research , 6 Verdun St , Nedlands , WA 6009 , Australia .
| | - Amy L Kretzmann
- School of Molecular Sciences , The University of Western Australia , 35 Stirling Hwy , Crawley , WA 6009 , Australia .
| | - Edina Wang
- Harry Perkins Institute of Medical Research , 6 Verdun St , Nedlands , WA 6009 , Australia .
| | - Diwei Ho
- School of Molecular Sciences , The University of Western Australia , 35 Stirling Hwy , Crawley , WA 6009 , Australia .
| | - Mark J Hackett
- Curtin Institute for Functional Molecules and Interfaces , Curtin Health Innovation Research Institute , Department of Chemistry , Curtin University , Bentley , WA 6845 , Australia
| | - Benjamin F Dessauvagie
- Anatomical Pathology, PathWest Laboratory Medicine , Fiona Stanley Hospital , Murdoch , WA , Australia.,School of Medicine , The University of Western Australia , Crawley , WA , Australia
| | - Nicole M Smith
- School of Molecular Sciences , The University of Western Australia , 35 Stirling Hwy , Crawley , WA 6009 , Australia .
| | - Andrew D Redfern
- School of Medicine , The University of Western Australia , Crawley , WA , Australia
| | - Charlene Waryah
- Harry Perkins Institute of Medical Research , 6 Verdun St , Nedlands , WA 6009 , Australia .
| | - Marck Norret
- School of Molecular Sciences , The University of Western Australia , 35 Stirling Hwy , Crawley , WA 6009 , Australia .
| | - K Swaminathan Iyer
- School of Molecular Sciences , The University of Western Australia , 35 Stirling Hwy , Crawley , WA 6009 , Australia .
| | - Pilar Blancafort
- Harry Perkins Institute of Medical Research , 6 Verdun St , Nedlands , WA 6009 , Australia .
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Acikalin D, Oner U, Can C, Acikalin MF, Colak E. Predictive Value of Maspin and Ki-67 Expression in Transurethral Resection Specimens in Patients with T1 Bladder Cancer. TUMORI JOURNAL 2018; 98:344-50. [DOI: 10.1177/030089161209800311] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022]
Abstract
Aims and background To evaluate the clinical significance of maspin and Ki-67 expression in patients with newly diagnosed T1 bladder cancer. Methods and study design Maspin and Ki-67 expression was investigated by immunohistochemistry from paraffin-embedded tissues of 68 patients undergoing transurethral resection for bladder cancer. Clinicopathological data were retrospectively reviewed from available charts and pathological reports. Maspin and Ki-67 expression levels were classified according to the staining percentage. Cases in which at least 5% of the tumor cells stained for maspin were scored as positive. Ki-67 labeling index was considered to be positive when samples demonstrated >10% reactivity. Results Maspin expression was found as an independent predictor of recurrence and progression (P <0.05). Patients with negative maspin expression were 2.191 times more likely to relapse than patients with positive maspin expression. Patients with negative maspin expression were 4.345 times more likely to progress than patients with positive maspin expression. Furthermore, the maspin-negative group was found to have shorter recurrence and progression-free survival (P <0.05). No significant association was found between maspin subcellular localization pattern and recurrence-free, progression-free or overall survival (P >0.05). There was no correlation between Ki-67 expression and tumor recurrence, progression or tumor-related death (P >0.05). Chi-square tests showed a significant relationship between Ki-67 expression and tumor size and tumor grade (P <0.05). Conclusions Our findings suggested that the evaluation of maspin expression in stage T1 bladder tumors is a useful prognostic marker for predicting the tumor behavior.
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Affiliation(s)
- Demet Acikalin
- Department of Pathology, Eskisehir
Osmangazi University Medical Faculty, Eskisehir, Turkey
| | - Ulku Oner
- Department of Pathology, Eskisehir
Osmangazi University Medical Faculty, Eskisehir, Turkey
| | - Cavit Can
- Department of Urology, Eskisehir
Osmangazi University Medical Faculty, Eskisehir, Turkey
| | - Mustafa F Acikalin
- Department of Pathology, Eskisehir
Osmangazi University Medical Faculty, Eskisehir, Turkey
| | - Ertugrul Colak
- Department of Biostatistics, Eskisehir
Osmangazi University Medical Faculty, Eskisehir, Turkey
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Thivyah Prabha A, Sekar D. Deciphering the molecular signaling pathways in breast cancer pathogenesis and their role in diagnostic and treatment modalities. GENE REPORTS 2017; 7:1-17. [DOI: 10.1016/j.genrep.2017.01.003] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/23/2022]
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Tamazato Longhi M, Magalhães M, Reina J, Morais Freitas V, Cella N. EGFR Signaling Regulates Maspin/SerpinB5 Phosphorylation and Nuclear Localization in Mammary Epithelial Cells. PLoS One 2016; 11:e0159856. [PMID: 27447178 PMCID: PMC4957797 DOI: 10.1371/journal.pone.0159856] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/07/2016] [Accepted: 07/08/2016] [Indexed: 12/13/2022] Open
Abstract
Maspin (SerpinB5) is a non-inhibitory serpin (serine protease inhibitor) with very diverse biological activities including regulation of cell adhesion, migration, death, control of gene expression and oxidative stress response. Initially described as a tumor and metastasis suppressor, clinical data brought controversies to the field, as some studies reported no correlation between SerpinB5 expression and prognosis value. These data underscore the importance of understanding SerpinB5 function in a normal physiological context and the molecular mechanism involved. Several SerpinB5 phosphoforms have been detected in different cell lines, but the signaling pathways involved and the biological significance of this post-translational modification in vivo remains to be explored. In this study we investigated SerpinB5 expression, subcellular localization and phosphorylation in different stages of the mouse mammary gland development and the signaling pathway involved. Here we show that SerpinB5 is first detected in late pregnancy, reaches its highest levels in lactation and remains at constant levels during post-lactational regression (involution). Using high resolution isoelectric focusing followed but immunoblot, we found at least 8 different phosphoforms of SerpinB5 during lactation, which decreases steadily at the onset of involution. In order to investigate the signaling pathway involved in SerpinB5 phosphorylation, we took advantage of the non-transformed MCF-10A model system, as we have previously observed SerpinB5 phosphorylation in these cells. We detected basal levels of SerpinB5 phosphorylation in serum- and growth factor-starved cells, which is due to amphiregulin autocrine activity on MCF-10A cells. EGF and TGF alpha, two other EGFR ligands, promote important SerpinB5 phosphorylation. Interestingly, EGF treatment is followed by SerpinB5 nuclear accumulation. Altogether, these data indicate that SerpinB5 expression and phosphorylation are developmentally regulated. In vitro analyses indicate that SerpinB5 phosphorylation is regulated by EGFR ligands, but EGF appears to be the only able to induce SerpinB5 nuclear localization.
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Affiliation(s)
- Mariana Tamazato Longhi
- Departamento de Biologia Celular e do Desenvolvimento, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil
| | - Magna Magalhães
- Departamento de Biologia Celular e do Desenvolvimento, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil
| | - Jeffrey Reina
- Departamento de Biologia Celular e do Desenvolvimento, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil
| | - Vanessa Morais Freitas
- Departamento de Biologia Celular e do Desenvolvimento, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil
| | - Nathalie Cella
- Departamento de Biologia Celular e do Desenvolvimento, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil
- * E-mail:
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Vereecken P, Reynaert S, Lalmand MC, Zouaoui-Boudjeltia K, Heenen M, Van Den Heule B, Petein M. Decreased Immunoreactive Maspin Expression in Intermediate Thickness and Thick Primary Melanoma Lesions. J Int Med Res 2016; 34:52-7. [PMID: 16604823 DOI: 10.1177/147323000603400106] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/15/2022] Open
Abstract
Maspin is a member of the serpin family of protease inhibitors. It is a 42 kDa cytoplasmic protein that is reported to have tumour suppressor activity. The loss of maspin gene expression is correlated with increased invasiveness and the risk of metastases in breast cancer. We studied maspin expression in primary melanoma lesions obtained from 76 patients. Immunostaining of 5 μm sections for maspin expression was obtained using the citrate antigen retrieval method. The extent of immunostaining was scored by recording the proportion of immunoreactive cells and the intensity of immunostaining. Our results demonstrated that maspin expression was down-regulated in intermediate thickness and thick melanoma lesions compared with thin lesions. These results suggest that loss of maspin expression might play a role in melanoma progression, invasion and metastatic dissemination. Further studies are needed to clarify the clinicopathological significance of maspin expression in melanoma.
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Affiliation(s)
- P Vereecken
- Department of Dermatology, Erasme University Hospital, Brussels, Belgium.
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Teoh SSY, Vieusseux J, Prakash M, Berkowicz S, Luu J, Bird CH, Law RHP, Rosado C, Price JT, Whisstock JC, Bird PI. Maspin is not required for embryonic development or tumour suppression. Nat Commun 2016; 5:3164. [PMID: 24445777 PMCID: PMC3905777 DOI: 10.1038/ncomms4164] [Citation(s) in RCA: 26] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/12/2013] [Accepted: 12/20/2013] [Indexed: 02/07/2023] Open
Abstract
Maspin (SERPINB5) is accepted as an important tumour suppressor lost in many cancers. Consistent with a critical role in development or differentiation maspin knockout mice die during early embryogenesis, yet clinical data conflict on the prognostic utility of maspin expression. Here to reconcile these findings we made conditional knockout mice. Surprisingly, maspin knockout embryos develop into overtly normal animals. Contrary to original reports, maspin re-expression does not inhibit tumour growth or metastasis in vivo, or influence cell migration, invasion or survival in vitro. Bioinformatic analyses reveal that maspin is not commonly under-expressed in cancer, and that perturbation of genes near maspin may in fact explain poor survival in certain patient cohorts with low maspin expression. A role for the serpin maspin has been described in both development and cancer. In this study, the authors demonstrate that maspin knockout mice develop normally and that maspin does not function as a tumour suppressor, suggesting that another gene at the maspin locus may be responsible for this activity.
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Affiliation(s)
- Sonia S Y Teoh
- Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Victoria 3800, Australia
| | - Jessica Vieusseux
- Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Victoria 3800, Australia
| | - Monica Prakash
- Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Victoria 3800, Australia
| | - Susan Berkowicz
- Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Victoria 3800, Australia
| | - Jennii Luu
- Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Victoria 3800, Australia
| | - Catherina H Bird
- Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Victoria 3800, Australia
| | - Ruby H P Law
- 1] Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Victoria 3800, Australia [2] Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics, School of Biomedical Sciences, Monash University, Victoria 3800, Australia
| | - Carlos Rosado
- 1] Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Victoria 3800, Australia [2] Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics, School of Biomedical Sciences, Monash University, Victoria 3800, Australia
| | - John T Price
- Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Victoria 3800, Australia
| | - James C Whisstock
- 1] Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Victoria 3800, Australia [2] Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics, School of Biomedical Sciences, Monash University, Victoria 3800, Australia
| | - Phillip I Bird
- Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Victoria 3800, Australia
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12
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Fortenberry Y. The role of serpins in tumor cell migration. Biol Chem 2015; 396:205-13. [PMID: 25381952 DOI: 10.1515/hsz-2014-0254] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/26/2014] [Accepted: 11/03/2014] [Indexed: 01/13/2023]
Abstract
Tumor cells are characterized by uncontrolled cell growth at a primary site that is caused by genetic alterations. Tumor cells that metastasize from their primary site to distant locations are commonly referred to as malignant. Cell migration is a critical step in this process. The ability of tumor cells to migrate and invade is partly controlled by proteolytic enzymes. These enzymes are secreted by either the tumor cells themselves or adjacent cells. They represent all classes of proteases, including serine and cysteine proteases. Serine proteases, in particular urokinase plasminogen activator (uPA), initiate a proteolytic cascade that culminates in degrading components of the extracellular matrix (ECM). Some serine proteases are controlled by a superfamily of proteins known as serpins. This minireview provides an overview of serpins that are vital in regulating tumor cell migration and progressing cancer.
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Farhanji B, Latifpour M, Alizadeh AM, Khodayari H, Khodayari S, Khaniki M, Ghasempour S. Tumor suppression effects of myoepithelial cells on mice breast cancer. Eur J Pharmacol 2015; 765:171-8. [PMID: 26297304 DOI: 10.1016/j.ejphar.2015.08.023] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/04/2015] [Revised: 08/12/2015] [Accepted: 08/17/2015] [Indexed: 01/20/2023]
Abstract
Several studies have assumed that myoepithelial cells (MECs) loss may contribute to epithelial tumor induction and/or progression. We adopted an in vitro assay and a syngeneic mice breast cancer model with histological and molecular characteristics resembling human lesions to evaluate tumor suppression effects of MECs. Flow cytometric, cell viability, blood chemistry, transmission electron microscope, immunohistochemistry and qRT-PCR assays were performed at the end of the study. We demonstrated that MECs could significantly suppress the viability of cancer cells at different time points (P<0.05). At the end of the fourth and fifth weeks, treated mice had smaller tumor volume compared with control animals. Average tumor volume was significantly less in treated groups than control group at days 21 (0.38±0.19 vs. 1.99±0.13 cm3), 28 (0.57±0.3 vs. 2.5±0.37 cm3) and 35 (0.7±0.35 vs. 2.65±0.4 cm3) after tumor cell injection (P<0.05). No hematological, hepatocellular, and renal toxicities were seen in MECs treated groups. Ultrastructural features revealed severe relationship between adjacent tumoral cells and loose interconnections of neoplastic cells in treated group. Immunohistochemical examinations of breast tumors showed high p63 and low alpha-smooth muscle actin protein expression in treated mice compared to control (P<0.05). MRNA expressions of TNF-α, smooth muscle-myosin heavy chain, connexin 43, and maspin were significantly up-regulated in breast tumor tissues in treated group compared to control (P<0.05). VEGF and alpha-smooth muscle actin mRNA expression were reduced in treated animals (P<0.05). The present study highlighted the potential tumor suppression effects of MECs on breast cancer in a typical animal model.
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Affiliation(s)
- Baharak Farhanji
- Iranian Tissue Bank & Research Center, Tehran University of Medical Sciences, Tehran, Iran
| | - Mostafa Latifpour
- Cancer Research Center, Tehran University of Medical Sciences, 1419733141 Tehran, Iran
| | - Ali Mohammad Alizadeh
- Cancer Research Center, Tehran University of Medical Sciences, 1419733141 Tehran, Iran.
| | - Hamid Khodayari
- Cancer Research Center, Tehran University of Medical Sciences, 1419733141 Tehran, Iran
| | - Saeed Khodayari
- Cancer Research Center, Tehran University of Medical Sciences, 1419733141 Tehran, Iran; Department of Pharmacology, Fasa University of Medical Sciences, Fasa, Iran
| | - Mahmood Khaniki
- Pathology Department, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran
| | - Sarieh Ghasempour
- Cancer Research Center, Tehran University of Medical Sciences, 1419733141 Tehran, Iran
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14
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Expression and localization of maspin in cervical cancer and its role in tumor progression and lymphangiogenesis. Arch Gynecol Obstet 2014; 289:373-82. [PMID: 23959090 PMCID: PMC3894428 DOI: 10.1007/s00404-013-2988-4] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/07/2013] [Accepted: 07/26/2013] [Indexed: 12/14/2022]
Abstract
Objectives Cervical cancer is the most common malignant tumor in female reproductive tract and primarily metastasizes through the lymphatic system that will affect prognosis of patients. Maspin, a member of the serine protease inhibitors (serpins) super family, has recently been indicated as a tumor suppressor in many cancers. In this study, we investigated the clinical significance of maspin expression, especially the subcellular location of maspin and its functional role in progression and lymphangiogenesis, in cervical squamous cell carcinoma. Methods Labelled streptavidin biotin method (LSAB) was used to determine cytoplasmic and nuclear maspin expressions, respectively, in 13 cases of normal cervix, 15 cases of cervical intraepithelial neoplasia grade 3 (CIN3), 62 cases of squamous cell carcinoma (SCC) of the uterine cervix, and 13 cases of pelvic lymphatic nodes which were all positive lymphatic nodes in our selected cancer cases. LSAB is also used to detect podoplanin which is used for counting density of lymphatic microvessels (LMVD). The clinical significance of subcellular maspin expression and the relationship between maspin expression and LMVD in cervical cancer are analyzed. Results Both cytoplasmic and nuclear maspin expressions in SCC were significantly weaker than those of normal cervix and CIN3. Nuclear maspin expression showed a peak in CIN3 and then dropped in SCC. Declined maspin expression was correlated with later clinical stage, increased LMVD, and lymphatic metastasis. Conclusions Our results suggest that subcellular location of maspin expression is a potential predictive factor in tumor progression and in patients’ prognosis of cervical cancer, and maspin plays a suppression role in lymphangiogenesis and metastasis.
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Liu Q, Qiao FY, Shi XW, Liu HY, Gong X, Wu YY. Promoter hypomethylation and increased maspin expression in preeclamptic placentas in a Chinese population. Placenta 2014; 35:876-82. [PMID: 25151033 DOI: 10.1016/j.placenta.2014.08.088] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 05/13/2014] [Revised: 07/15/2014] [Accepted: 08/05/2014] [Indexed: 12/14/2022]
Abstract
OBJECTIVE Preeclampsia is thought to begin with shallow trophoblast invasion and inadequate spiral artery remodeling. Maspin, a tumor-suppressor gene, plays a regulatory role in trophoblast invasion and motility. The tissue-specific methylation of the maspin promoter can regulate maspin gene expression in various cancers. We sought to detect maspin gene expression and assess the degrees of methylation of maspin promoter regions in preeclamptic placentas in the Han Chinese population and to investigate the potential role of maspin in the pathophysiology of preeclampsia. METHODS We conducted RT-PCR, immunohistochemistry and western blotting to characterize maspin gene expression and protein levels in the placentas from normal and preeclamptic pregnancies. Finally, using methylation-specific PCR and bisulfite sequencing PCR, we detected the degrees of methylation of the promoter regions of maspin in each of the two studied groups. RESULTS Maspin expression was increased at the mRNA and protein levels in the preeclamptic placentas compared to the control group. Maspin immunohistochemical staining revealed positive staining in the syncytio-cytotrophoblast layers and more diffuse staining in the preeclamptic group. The mean methylation level of the analyzed promoter region was significantly hypomethylated in the preeclamptic placentas compared to the control placentas, pointing to a negative relationship between maspin promoter methylation and gene expression. DISCUSSION Hypomethylation of the maspin promoter results in increased expression of maspin in preeclamptic placentas, which suggests a negative relationship between maspin methylation and maspin expression in this Han Chinese population. Thus, maspin is likely involved in the etiology of preeclampsia.
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Affiliation(s)
- Q Liu
- Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China
| | - F Y Qiao
- Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China
| | - X W Shi
- Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China
| | - H Y Liu
- Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China
| | - X Gong
- Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China
| | - Y Y Wu
- Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
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Jacobo-Herrera NJ, Pérez-Plasencia C, Camacho-Zavala E, González GF, Urrutia EL, García-Castillo V, Zentella-Dehesa A. Clinical evidence of the relationship between aspirin and breast cancer risk (review). Oncol Rep 2014; 32:451. [PMID: 24927467 DOI: 10.3892/or.2014.3270] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/21/2014] [Accepted: 05/15/2014] [Indexed: 11/06/2022] Open
Abstract
In the search for new therapeutic alternatives against cancer, either as a preventive treatment or for advanced stages, it is common to appeal to well-known drugs used for the treatment of other diseases that may interfere with the metabolic pathways involved in carcinogenesis. Non-steroidal anti-inflammatory drugs (NSAIDs) display anticancer activity through the inhibition of the COX-2 enzyme, triggering processes such as apoptosis, a reduction in proliferation and inhibition of carcinogenesis. Breast cancer is a neoplasm with the highest incidence and mortality rate among young women worldwide. Epidemiologic data have shown that drugs such as NSAIDs, particularly aspirin, reduce the relative risk of breast cancer. However, in the subgroup of responsive patients, dose, time and frequency of use have not yet been established. Here, we review the reports published during the last 10 years regarding the relationship between breast cancer and aspirin.
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Affiliation(s)
- Nadia J Jacobo-Herrera
- Unidad de Bioquímica, Instituto Nacional de Ciencias Médicas y Nutrición 'Salvador Zubirán', Tlalpan 14000, Mexico, D.F., Mexico
| | - Carlos Pérez-Plasencia
- Unidad de Biomedicina FES-Iztacala, Universidad Nacional Autónoma de México UNAM, Tlalnepantla 54090, Mexico
| | - Elizabeth Camacho-Zavala
- Unidad de Bioquímica, Instituto Nacional de Ciencias Médicas y Nutrición 'Salvador Zubirán', Tlalpan 14000, Mexico, D.F., Mexico
| | - Gabriela Figueroa González
- Unidad de Biomedicina FES-Iztacala, Universidad Nacional Autónoma de México UNAM, Tlalnepantla 54090, Mexico
| | - Eduardo López Urrutia
- Laboratorio de Oncogenómica, Instituto Nacional de Cancerología, Tlalpan 14080, Mexico, D.F., Mexico
| | - Verónica García-Castillo
- Unidad de Biomedicina FES-Iztacala, Universidad Nacional Autónoma de México UNAM, Tlalnepantla 54090, Mexico
| | - Alejandro Zentella-Dehesa
- Unidad de Bioquímica, Instituto Nacional de Ciencias Médicas y Nutrición 'Salvador Zubirán', Tlalpan 14000, Mexico, D.F., Mexico
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Maspin expression and melanoma progression: a matter of sub-cellular localization. Mod Pathol 2014; 27:412-9. [PMID: 24030740 DOI: 10.1038/modpathol.2013.157] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/22/2013] [Revised: 07/05/2013] [Accepted: 07/07/2013] [Indexed: 02/06/2023]
Abstract
Maspin, a member of the serpin family of protease inhibitors, is involved in key processes of cancer progression. Its biological activity seems to be cancer and compartment specific, with the protein acting either as a suppressor or as a tumor promoter in different cancer types. Characterization of maspin expression and its sub-cellular localization in melanoma is missing, hence, we aim to investigate its possible association with melanoma prognostic factors and disease progression. Nuclear and cytoplasmic maspin expression were evaluated on 60 nevi, 152 primary lesions, and 106 melanoma metastases using tissue microarrays and immunohistochemistry. The association between maspin immunoreactivity and patient's clinic-pathological features was evaluated. Multivariate logistic models and survival analyses were performed for maspin expression in primary melanomas. Nuclear maspin was detected in 8% nevi, 49% primary melanomas, and 28% metastases, whereas cytoplasmic maspin in 12% nevi, 18% primary lesions, and 9% metastases. In univariate analysis, nuclear maspin expression in primary melanomas was significantly associated with melanoma prognostic factors (nodular histotype, tumor thickness, mitotic rate, and ulceration) and disease stage, whereas cytoplasmic maspin was observed at higher frequency in thin superficial spreading melanomas, without mitosis. In multivariate analysis, nuclear maspin remained significantly associated with risk of developing a tumor prone to disease progression and, accordingly, with significantly shorter disease-free and overall survival. In this study, maspin was expressed at highest frequency in primary lesions and when expressed in the nuclei, was significantly associated with poor prognostic markers, melanoma recurrence, and worse survival. The present study suggests a tumor-suppressive effect of cytoplasmic maspin and a tumor-promoting effect of nuclear maspin, which open the discussion on its potential use in cancer therapy.
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Machowska M, Wachowicz K, Sopel M, Rzepecki R. Nuclear location of tumor suppressor protein maspin inhibits proliferation of breast cancer cells without affecting proliferation of normal epithelial cells. BMC Cancer 2014; 14:142. [PMID: 24581141 PMCID: PMC3975902 DOI: 10.1186/1471-2407-14-142] [Citation(s) in RCA: 35] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/22/2013] [Accepted: 02/11/2014] [Indexed: 12/13/2022] Open
Abstract
Background Maspin, which is classified as a tumor suppressor protein, is downregulated in many types of cancer. Several studies have suggested potential anti-proliferative activity of maspin as well as sensitizing activity of maspin for therapeutic cytotoxic agents in breast cancer tissue culture and animal models. All of the experimental data gathered so far have been based on studies with maspin localized cytoplasmically, while maspin in breast cancer tumor cells may be located in the cytoplasm, nucleus or both. In this study, the effect of maspin cytoplasmic and nuclear location and expression level on breast cancer proliferation and patient survival was studied. Methods Tissue sections from 166 patients with invasive ductal breast cancer were stained by immunohistochemistry for maspin and Ki-67 protein. The localization and expression level of maspin were correlated with estimated patient overall survival and percent of Ki-67-positive cells. In further studies, we created constructs for transient transfection of maspin into breast cancer cells with targeted cytoplasmic and nuclear location. We analyzed the effect of maspin location in normal epithelial cell line MCF10A and three breast cancer cell lines - MCF-7, MDA-MB-231 and SKBR-3 - by immunofluorescence and proliferation assay. Results We observed a strong positive correlation between moderate and high nuclear maspin level and survival of patients. Moreover, a statistically significant negative relationship was observed between nuclear maspin and Ki-67 expression in patients with invasive ductal breast cancer. Spearman’s correlation analysis showed a negative correlation between level of maspin localized in nucleus and percentage of Ki-67 positive cells. No such differences were observed in cells with cytoplasmic maspin. We found a strong correlation between nuclear maspin and loss of Ki-67 protein in breast cancer cell lines, while there was no effect in normal epithelial cells from breast. The anti-proliferative effect of nuclear maspin on breast cancer cells was statistically significant in comparison to cytoplasmic maspin. Conclusions Our results suggest that nuclear maspin localization may be a prognostic factor in breast cancer and may have a strong therapeutic potential in gene therapy. Moreover, these data provide a new insight into the role of cytoplasmic and nuclear fractions of maspin in breast cancer.
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Affiliation(s)
| | | | | | - Ryszard Rzepecki
- Laboratory of Nuclear Proteins, Faculty of Biotechnology, University of Wroclaw, 63/77 Przybyszewskiego Street, 51-148 Wrocław, Poland.
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Hayashi M, Jono H, Shinriki S, Nakamura T, Guo J, Sueta A, Tomiguchi M, Fujiwara S, Yamamoto-Ibusuki M, Murakami KI, Yamashita S, Yamamoto Y, Li JD, Iwase H, Ando Y. Clinical significance of CYLD downregulation in breast cancer. Breast Cancer Res Treat 2014; 143:447-57. [DOI: 10.1007/s10549-013-2824-3] [Citation(s) in RCA: 58] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/09/2013] [Accepted: 12/23/2013] [Indexed: 12/01/2022]
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Siddiqui RA, Harvey KA, Walker C, Altenburg J, Xu Z, Terry C, Camarillo I, Jones-Hall Y, Mariash C. Characterization of synergistic anti-cancer effects of docosahexaenoic acid and curcumin on DMBA-induced mammary tumorigenesis in mice. BMC Cancer 2013; 13:418. [PMID: 24034496 PMCID: PMC3848456 DOI: 10.1186/1471-2407-13-418] [Citation(s) in RCA: 45] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/02/2013] [Accepted: 09/09/2013] [Indexed: 01/09/2023] Open
Abstract
BACKGROUND The major obstacles to the successful use of individual nutritional compounds as preventive or therapeutic agents are their efficacy and bioavailability. One approach to overcoming this problem is to use combinations of nutrients to induce synergistic effects. The objective of this research was to investigate the synergistic effects of two dietary components: docosahexaenoic acid (DHA), an omega-3 fatty acid present in cold-water fish, and curcumin (CCM), an herbal nutrient present in turmeric, in an in vivo model of DMBA-induced mammary tumorigenesis in mice. METHODS We used the carcinogen DMBA to induce breast tumors in SENCAR mice on control, CCM, DHA, or DHA + CCM diets. Appearance and tumor progression were monitored daily. The tumors were harvested 15 days following their first appearance for morphological and immunohistological analysis. Western analysis was performed to determine expression of maspin and survivin in the tumor tissues. Characterization of tumor growth was analyzed using appropriate statistical methods. Otherwise all other results are reported as mean ± SD and analyzed with one-way ANOVA and Tukey's post hoc procedure. RESULTS Analysis of gene microarray data indicates that combined treatment with DHA + CCM altered the profile of "PAM50" genes in the SK-BR-3 cell line from an ER⁻/Her-2⁺ to that resembling a "normal-like" phenotype. The in vivo studies demonstrated that DHA + CCM treatment reduced the incidence of breast tumors, delayed tumor initiation, and reduced progression of tumor growth. Dietary treatment had no effect on breast size development, but tumors from mice on a control diet (untreated) were less differentiated than tumors from mice fed CCM or DHA + CCM diets. The synergistic effects also led to increased expression of the pro-apoptotic protein, maspin, but reduced expression of the anti-apoptotic protein, survivin. CONCLUSIONS The SK-BR-3 cells and DMBA-induced tumors, both with an ER⁻ and Her-2⁺ phenotype, were affected by the synergistic interaction of DHA and CCM. This suggests that the specific breast cancer phenotype is an important factor for predicting efficacy of these nutraceuticals. The combination of DHA and CCM is potentially a dietary supplemental treatment for some breast cancers, likely dependent upon the molecular phenotype of the cancer.
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Affiliation(s)
- Rafat A Siddiqui
- Cellular Biochemistry Laboratory, Indiana University Health, Indianapolis, IN 46202, USA.
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Stark AM, Schem C, Maass N, Hugo HH, Jonat W, Mehdorn HM, Held-Feindt J. Expression of metastasis suppressor gene maspin is reduced in breast cancer brain metastases and correlates with the estrogen receptor status. Neurol Res 2013; 32:303-8. [DOI: 10.1179/016164109x12518779082192] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/07/2023]
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Pföhler C, Knöpflen T, Körner R, Vogt T, Rösch A, Müller CSL. Maspin expression in the invasive margin of primary melanomas may reflect an aggressive tumor phenotype. J Dtsch Dermatol Ges 2013; 11:993-9. [PMID: 23848940 DOI: 10.1111/ddg.12121] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/17/2013] [Accepted: 04/01/2013] [Indexed: 12/13/2022]
Abstract
BACKGROUND The role of maspin has been discussed controversially in different tumors. In the majority of malignant tumors, maspin seems to act as a tumor suppressor. However, data about maspin expression as well as its function in melanoma are very inconsistent. OBJECTIVE To investigate the expression of maspin in melanomas and to correlate the intensity of maspin staining with prognostic parameters of the tumor and with progression-free and overall survival. PATIENTS AND METHODS Primary melanomas from 47 patients were investigated for maspin expression using immunohistochemistry. RESULTS Maspin was heterogeneously expressed predominantly in the cytoplasm of melanoma cells. Maspin staining intensity in the invasive part of the tumor correlated with parameters of prognosis such as Clark level (p = 0.05), tumor thickness (p = 0.002) and stage of disease (p = 0.023). Maspin staining intensity in the invasive front of the tumor significantly correlated with death from disease (p = 0.007) and shortened overall survival (p = 0.007). CONCLUSIONS In accordance with data concerning maspin expression in colorectal cancers, the expression of this protein in the invasive front of primary melanomas seems to correlate with local infiltration and tumor aggressiveness. Strong maspin expression in the invasive margin of primary melanomas therefore might reflect an aggressive tumor phenotype.
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Affiliation(s)
- Claudia Pföhler
- Saarland University Hospital, Department of Dermatology, Homburg/Saar, Germany
| | - Tobias Knöpflen
- Saarland University Hospital, Department of Dermatology, Homburg/Saar, Germany
| | - Rebecca Körner
- Saarland University Hospital, Department of Dermatology, Homburg/Saar, Germany
| | - Thomas Vogt
- Saarland University Hospital, Department of Dermatology, Homburg/Saar, Germany
| | - Alexander Rösch
- Saarland University Hospital, Department of Dermatology, Homburg/Saar, Germany
| | - Cornelia S L Müller
- Saarland University Hospital, Department of Dermatology, Homburg/Saar, Germany
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Bodenstine TM, Seftor REB, Khalkhali-Ellis Z, Seftor EA, Pemberton PA, Hendrix MJC. Maspin: molecular mechanisms and therapeutic implications. Cancer Metastasis Rev 2013; 31:529-51. [PMID: 22752408 DOI: 10.1007/s10555-012-9361-0] [Citation(s) in RCA: 81] [Impact Index Per Article: 6.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/12/2022]
Abstract
Maspin, a non-inhibitory member of the serine protease inhibitor superfamily, has been characterized as a tumor suppressor gene in multiple cancer types. Among the established anti-tumor effects of Maspin are the inhibition of cancer cell invasion, attachment to extracellular matrices, increased sensitivity to apoptosis, and inhibition of angiogenesis. However, while significant experimental data support the role of Maspin as a tumor suppressor, clinical data regarding the prognostic implications of Maspin expression have led to conflicting results. This highlights the need for a better understanding of the context dependencies of Maspin in normal biology and how these are perturbed in the context of cancer. In this review, we outline the regulation and roles of Maspin in normal and developmental biology while discussing novel evidence and emerging theories related to its functions in cancer. We provide insight into the immense therapeutic potential of Maspin and the challenges related to its successful clinical translation.
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Affiliation(s)
- Thomas M Bodenstine
- Children's Hospital of Chicago Research Center, Robert H. Lurie Comprehensive Cancer Center, Northwestern University Feinberg School of Medicine, 225 E. Chicago Avenue, Box 222, Chicago, IL 60611, USA
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Berardi R, Morgese F, Onofri A, Mazzanti P, Pistelli M, Ballatore Z, Savini A, De Lisa M, Caramanti M, Rinaldi S, Pagliaretta S, Santoni M, Pierantoni C, Cascinu S. Role of maspin in cancer. Clin Transl Med 2013; 2:8. [PMID: 23497644 PMCID: PMC3602294 DOI: 10.1186/2001-1326-2-8] [Citation(s) in RCA: 88] [Impact Index Per Article: 7.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/29/2012] [Accepted: 01/28/2013] [Indexed: 02/08/2023] Open
Abstract
Maspin (mammary serine protease inhibitor), is a member of the serine protease inhibitor/non-inhibitor superfamily. Its expression is down-regulated in breast, prostate, gastric and melanoma cancers but over-expressed in pancreatic, gallbladder, colorectal, and thyroid cancers suggesting that maspin may play different activities in different cell types. However, maspin expression seems to be correlated with better prognosis in prostate, bladder, lung, gastric, colorectal, head and neck, thyroid and melanoma cancer. In breast and ovarian cancer maspin significance is associated with its subcellular localization: nucleus maspin expression correlates with a good prognosis, whilst in pancreatic cancer it predicts a poor prognosis. Since tumor metastasis requires the detachment and invasion of tumor cells through the basement membrane and stroma, a selectively increased adhesion by the presence of maspin may contribute to the inhibition of tumor metastasis. Furthermore the different position of maspin inside the cell or its epigenetic modifications may explain the different behavior of the expression of maspin between tumors. The expression of maspin might be useful as a prognostic and possibly predictive factor for patients with particular types of cancer and data can guide physicians in selecting therapy. Its expression in circulating tumor cells especially in breast cancer, could be also useful in clinical practice along with other factors, such as age, comorbidities, blood examinations in order to select the best therapy to be carried out. Focusing on the malignancies in which maspin showed a positive prognostic value, therapeutic approaches studied so far aimed to re-activate a dormant tumor suppressor gene by designed transcription factors, to hit the system that inhibits the expression of maspin, to identify natural substances that can determine the activation and the expression of maspin or possible "molecules binds" to introduce maspin in cancer cell and gene therapy capable of up-regulating the maspin in an attempt to reduce primarily the risk of metastasis.Further studies in these directions are necessary to better define the therapeutic implication of maspin.
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Affiliation(s)
- Rossana Berardi
- Medical Oncology Unit, Università Politecnica delle Marche, Azienda Ospedaliero-Universitaria Ospedali Riuniti Umberto I, GM Lancisi, G Salesi di Ancona, Via Conca, Ancona, 71-60126, Italy
| | - Francesca Morgese
- Medical Oncology Unit, Università Politecnica delle Marche, Azienda Ospedaliero-Universitaria Ospedali Riuniti Umberto I, GM Lancisi, G Salesi di Ancona, Via Conca, Ancona, 71-60126, Italy
| | - Azzurra Onofri
- Medical Oncology Unit, Università Politecnica delle Marche, Azienda Ospedaliero-Universitaria Ospedali Riuniti Umberto I, GM Lancisi, G Salesi di Ancona, Via Conca, Ancona, 71-60126, Italy
| | - Paola Mazzanti
- Medical Oncology Unit, Università Politecnica delle Marche, Azienda Ospedaliero-Universitaria Ospedali Riuniti Umberto I, GM Lancisi, G Salesi di Ancona, Via Conca, Ancona, 71-60126, Italy
| | - Mirco Pistelli
- Medical Oncology Unit, Università Politecnica delle Marche, Azienda Ospedaliero-Universitaria Ospedali Riuniti Umberto I, GM Lancisi, G Salesi di Ancona, Via Conca, Ancona, 71-60126, Italy
| | - Zelmira Ballatore
- Medical Oncology Unit, Università Politecnica delle Marche, Azienda Ospedaliero-Universitaria Ospedali Riuniti Umberto I, GM Lancisi, G Salesi di Ancona, Via Conca, Ancona, 71-60126, Italy
| | - Agnese Savini
- Medical Oncology Unit, Università Politecnica delle Marche, Azienda Ospedaliero-Universitaria Ospedali Riuniti Umberto I, GM Lancisi, G Salesi di Ancona, Via Conca, Ancona, 71-60126, Italy
| | - Mariagrazia De Lisa
- Medical Oncology Unit, Università Politecnica delle Marche, Azienda Ospedaliero-Universitaria Ospedali Riuniti Umberto I, GM Lancisi, G Salesi di Ancona, Via Conca, Ancona, 71-60126, Italy
| | - Miriam Caramanti
- Medical Oncology Unit, Università Politecnica delle Marche, Azienda Ospedaliero-Universitaria Ospedali Riuniti Umberto I, GM Lancisi, G Salesi di Ancona, Via Conca, Ancona, 71-60126, Italy
| | - Silvia Rinaldi
- Medical Oncology Unit, Università Politecnica delle Marche, Azienda Ospedaliero-Universitaria Ospedali Riuniti Umberto I, GM Lancisi, G Salesi di Ancona, Via Conca, Ancona, 71-60126, Italy
| | - Silvia Pagliaretta
- Medical Oncology Unit, Università Politecnica delle Marche, Azienda Ospedaliero-Universitaria Ospedali Riuniti Umberto I, GM Lancisi, G Salesi di Ancona, Via Conca, Ancona, 71-60126, Italy
| | - Matteo Santoni
- Medical Oncology Unit, Università Politecnica delle Marche, Azienda Ospedaliero-Universitaria Ospedali Riuniti Umberto I, GM Lancisi, G Salesi di Ancona, Via Conca, Ancona, 71-60126, Italy
| | - Chiara Pierantoni
- Medical Oncology Unit, Università Politecnica delle Marche, Azienda Ospedaliero-Universitaria Ospedali Riuniti Umberto I, GM Lancisi, G Salesi di Ancona, Via Conca, Ancona, 71-60126, Italy
| | - Stefano Cascinu
- Medical Oncology Unit, Università Politecnica delle Marche, Azienda Ospedaliero-Universitaria Ospedali Riuniti Umberto I, GM Lancisi, G Salesi di Ancona, Via Conca, Ancona, 71-60126, Italy
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Kim M, Ju H, Lim B, Kang C. Maspin genetically and functionally associates with gastric cancer by regulating cell cycle progression. Carcinogenesis 2012; 33:2344-50. [DOI: 10.1093/carcin/bgs280] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/29/2023] Open
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Goulet B, Chan G, Chambers AF, Lewis JD. An emerging role for the nuclear localization of maspin in the suppression of tumor progression and metastasis. Biochem Cell Biol 2011; 90:22-38. [PMID: 22047058 DOI: 10.1139/o11-053] [Citation(s) in RCA: 31] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
Maspin, a member of the serpin family of serine protease inhibitors, was originally identified as a tumor suppressor that is expressed in normal mammary epithelial cells but is reduced or absent in breast carcinomas. Early enthusiasm for maspin as a biomarker for disease progression has been tempered by clinical data that associates maspin with favourable outcomes in some studies and poor prognosis in others. Here, we review all of the published clinical studies for maspin in breast and ovarian cancers and propose that the apparent discordance between clinical reports is a consequence of differential cellular distribution of maspin. Indeed, it was thought that an extracellular pool of maspin possessed tumor suppressor activity, acting by inhibiting migration and increasing cell adhesion. Recent evidence from our group and others indicates, however, that the nuclear localization of maspin in cancer cells is necessary for its tumor suppressor activity. We provide additional data here to demonstrate that nuclear-localized maspin binds to chromatin and is required to effectively prevent cells from metastasizing. Our knowledge of other serpins that localize to the nucleus should help to inform future studies of nuclear maspin. Elucidation of the molecular mechanisms regulating the localization and activities of maspin should pave the way for the development of improved diagnostics and therapies for cancer.
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Affiliation(s)
- Brigitte Goulet
- London Regional Cancer Program, Translational Prostate Cancer Research Group, London, ON N6A 4L6, Canada
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27
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Ghaedi M, Soleimani M, Taghvaie NM, Sheikhfatollahi M, Azadmanesh K, Lotfi AS, Wu J. Mesenchymal stem cells as vehicles for targeted delivery of anti-angiogenic protein to solid tumors. J Gene Med 2011; 13:171-80. [PMID: 21449040 DOI: 10.1002/jgm.1552] [Citation(s) in RCA: 31] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/17/2022] Open
Abstract
BACKGROUND Inhibition of tumor-induced angiogenesis may restrict tumor growth and metastasis. Long-term systemic delivery of angiogenic inhibitors is associated with toxicity, as well as other severe side-effects. The utility of cells as vehicles for gene therapy to deliver therapeutic molecules has been suggested to represent an efficient approach. Mesenchymal stem cells (MSCs) exhibit a tropism to cancer tissue, and may serve as a cellular delivery vehicle and a local producer of anti-angiogenic agents. METHODS In the present study, we attempted to assess production of the transgene, α1-antitrypsin (AAT), in lentivirus-transduced human MSCs and its cytotoxicity against human umbilical cord vein endothelial cells (HUVEC). The secreted protein from these effector cells was determined by an enzyme-linked immunosorbent assay. The cytotoxicity of hMSCs that overexpress the human AAT gene against HUVEC was evaluated with an apoptotic assay. RESULTS Lentivirus-transduced hMSCs produced functional AAT and displayed much higher cytotoxicity against HUVEC than untransduced hMSCs. Moreover, AAT secreted from transduced hMSCs significantly inhibited HUVEC proliferation compared to untransduced hMSCs. The data obtained demonstrate for the first time that genetically modified hMSCs released abundant and functional AAT that caused obvious cytotoxicity to HUVEC. CONCLUSIONS hMSC may serve as an effective platform for the targeted delivery of therapeutic proteins to cancer sites.
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Affiliation(s)
- Mahboobe Ghaedi
- Department of Internal Medicine, Transplant Research Program, University of California, Davis Medical Center, Sacramento, CA 95817, USA
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28
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Goulet B, Kennette W, Ablack A, Postenka CO, Hague MN, Mymryk JS, Tuck AB, Giguère V, Chambers AF, Lewis JD. Nuclear localization of maspin is essential for its inhibition of tumor growth and metastasis. J Transl Med 2011; 91:1181-7. [PMID: 21502940 DOI: 10.1038/labinvest.2011.66] [Citation(s) in RCA: 45] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/19/2023] Open
Abstract
Maspin (mammary serine protease inhibitor or SerpinB5) acts as a tumor suppressor when overexpressed in aggressive cancer cell lines. However, its role in human cancer is controversial. Maspin expression has been associated with a poor prognosis in some studies, whereas in others, with favorable outcome. The clinical data suggest, however, that nuclear-localized maspin is associated with improved survival. We hypothesized that the tumor suppressor activity of maspin may require nuclear localization, and that the discordance between clinical and experimental reports is a consequence of the variable subcellular distribution of maspin. Furthermore, we surmized that nuclear maspin could function as a tumor suppressor through the regulation of genes involved in tumor growth and invasion. Maspin or maspin fused to a nuclear export signal were expressed in metastatic human breast and epidermoid carcinoma cell lines. We found that pan-cellular localized maspin inhibited in vivo tumor growth and metastasis when assessed in xenograft chicken embryo and murine mammary fat pad injection models. However, when maspin was excluded from the nucleus via a nuclear exclusion signal, it no longer functioned as a metastasis suppressor. Using chromatin immunoprecipitation, we show that nuclear maspin was enriched at the promoter of colony-stimulating factor-1 (CSF-1) and associated with diminished levels of CSF-1 mRNA. Our findings demonstrate that the nuclear localization of maspin is required for its tumor and metastasis suppressor functions in vivo, and suggest that its mechanism of action involves, in part, direct association of maspin with target genes.
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29
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Villares GJ, Zigler M, Bar-Eli M. The emerging role of the thrombin receptor (PAR-1) in melanoma metastasis--a possible therapeutic target. Oncotarget 2011; 2:8-17. [PMID: 21378407 PMCID: PMC3248147 DOI: 10.18632/oncotarget.211] [Citation(s) in RCA: 39] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/25/2022] Open
Abstract
Melanoma remains as the deadliest form of skin cancer with limited and inefficient treatment options available for patients with metastatic disease. Within the last decade, the thrombin receptor, Protease Activated Receptor-1, has been described as an essential gene involved in the progression of human melanoma. PAR-1 is known to activate adhesive, invasive and angiogenic factors to promote melanoma metastasis. It is overexpressed not only in metastatic melanoma cell lines but is also highly expressed in metastatic lesions as compared to primary nevi and normal skin. Recently, PAR-1 has been described to regulate the gap junction protein Connexin 43 and the tumor suppressor gene Maspin to promote the metastatic melanoma phenotype. Herein, we review the role of PAR-1 in the progression of melanoma as well as utilizing PAR-1-regulated genes as potential therapeutic targets for melanoma treatment.
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Affiliation(s)
- Gabriel J Villares
- The University of Texas, MD Anderson Cancer Center, 1515 Holcombe Blvd, Unit 173 Houston, TX, USA
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Mauerer A, Roesch A, Hafner C, Stempfl T, Wild P, Meyer S, Landthaler M, Vogt T. Identification of new genes associated with melanoma. Exp Dermatol 2011; 20:502-7. [PMID: 21410771 DOI: 10.1111/j.1600-0625.2011.01254.x] [Citation(s) in RCA: 36] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/18/2022]
Abstract
PURPOSE Repeated failures in melanoma therapy made clear that the molecular mechanisms leading to melanoma are still poorly understood. In this study, we aim to provide a more comprehensive understanding of the transcriptional profiles and signalling pathways associated with melanoma. METHODS Gene expression was analysed using the Affymetrix Human Genome U133A 2.0 GeneChip arrays. To avoid culture artifacts, we used microdissected fresh frozen material of 18 melanocytic nevi (MN), 20 primary melanomas (PM) and 20 metastatic melanomas (MM). Statistical analysis was performed with Genomatix Chipinspector, Ingenuity™ Software, SPSS Software and Partek Genomic Suite 6.4. Expression levels of selected transcripts were verified by quantitative real-time RT-PCR and immunostaining of a tissue microarray sampling more than 280 cases of MN, PM and MM with known clinical outcome. RESULTS A total of 284 differentially expressed genes was detected in PM compared with MN and 189 genes in MM compared with PM affecting common cancer pathways such as MAPK-, Wnt- and Notch-signalling. Using principal component analysis, the samples could be grouped according to their histological entity. We identified a panel of novel melanoma-associated markers: frizzled-related protein, an antagonist of Wnt; tranducin-like enhancer of split 1, a transcription factor partner of TCF/LEF-1; CNTN1, an activator of Notch signalling; two Serpin peptidase inhibitors, Serpin B3/B4 and the TGF-β family member GDF15, the latter with association to MAPK-signalling.
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Affiliation(s)
- Andreas Mauerer
- Department of Dermatology, University of Regensburg, Regensburg, Germany
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31
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Sahin M, Sahin E, Gümüşlü S, Erdoğan A, Gültekin M. DNA methylation or histone modification status in metastasis and angiogenesis-related genes: a new hypothesis on usage of DNMT inhibitors and S-adenosylmethionine for genome stability. Cancer Metastasis Rev 2011; 29:655-76. [PMID: 20821252 DOI: 10.1007/s10555-010-9253-0] [Citation(s) in RCA: 31] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/29/2023]
Abstract
Metastasis is a leading cause of mortality and morbidity in cancer. This process needs angiogenesis. The biology underlying cancer, metastasis, and angiogenesis has been investigated so as to determine the therapeutic targets. Invasive and metastatic cancer cells have undergone numerous genetic and epigenetic changes, manifested by cytoskeletal changes, loss of adhesion, and expression of proteolytic enzymes that degrade the basement membrane. Additionally, in endothelial cells, some epigenetic modifications occur during the formation of angiogenesis. Researchers have used some methylation inhibitors, histone deacetylase inhibitors, or methylating agents (such as S-adenosylmethionine, SAM) against cancer and angiogenesis. Although they are effective to beat these diseases, each one results in differentiation or changes in genome structure. We review epigenetically modified genes related with angiogenesis and metastasis in cancer and endothelial cells, and suggest a new proposal. This hypothesis has discussed the importance of the usage of DNA methylation inhibitors together with SAM to prevent tumor progression and genome instability or changes resulting in additional diseases.
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Affiliation(s)
- Mehmet Sahin
- Health Sciences Research Centre, Faculty of Medicine, Akdeniz University, 07070 Antalya, Turkey.
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32
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Beltran AS, Blancafort P. Reactivation of MASPIN in non-small cell lung carcinoma (NSCLC) cells by artificial transcription factors (ATFs). Epigenetics 2011; 6:224-35. [PMID: 20948306 DOI: 10.4161/epi.6.2.13700] [Citation(s) in RCA: 41] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/06/2023] Open
Abstract
Tumor suppressor genes have antiproliferative and antimetastatic functions, and thus, they negatively affect tumor progression. Reactivating specific tumor suppressor genes would offer an important therapeutic strategy to block tumor progression. Mammary Serine Protease Inhibitor (MASPIN) is a tumor suppressor gene that is not mutated or rearranged in tumor cells, but is silenced during metastatic progression by transcriptional and epigenetic mechanisms. In this work, we have investigated the ability of Artificial Transcription Factors (ATFs) to reactivate MASPIN expression and to reduce tumor growth and metastatic dissemination in Non-Small Cell Lung Carcinoma (NSCLC) cell lines carrying a hypermethylated MASPIN promoter. We found that the ATFs linked to transactivator domains were able to demethylate the MASPIN promoter. Consistently, we observed that co-treatment of ATF-transduced cells with methyltransferase inhibitors enhanced MASPIN expression as well as induction of tumor cell apoptosis. In addition to tumor suppressive functions, restoration of endogenous MASPIN expression was accompanied by inhibition of metastatic dissemination in nude mice. ATF-mediated reactivation of MASPIN lead to changes in cell motility and to induction of E-CADHERIN. These data suggest that ATFs are able to reprogram aggressive lung tumor cells towards a more epithelial, differentiated phenotype, and thus, represent novel therapeutic agents for metastatic lung cancers.
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Affiliation(s)
- Adriana S Beltran
- Department of Pharmacology, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA
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Protease activated receptor-1 inhibits the Maspin tumor-suppressor gene to determine the melanoma metastatic phenotype. Proc Natl Acad Sci U S A 2010; 108:626-31. [PMID: 21187389 DOI: 10.1073/pnas.1006886108] [Citation(s) in RCA: 38] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/14/2022] Open
Abstract
The thrombin receptor protease activated receptor-1 (PAR-1) is overexpressed in metastatic melanoma cell lines and tumor specimens. Previously, we demonstrated a significant reduction in tumor growth and experimental lung metastasis after PAR-1 silencing via systemic delivery of siRNA encapsulated into nanoliposomes. Gene expression profiling identified a 40-fold increase in expression of Maspin in PAR-1-silenced metastatic melanoma cell lines. Maspin promoter activity was significantly increased after PAR-1 silencing, suggesting that PAR1 negatively regulates Maspin at the transcriptional level. ChIP analyses revealed that PAR-1 decreases binding of Ets-1 and c-Jun transcription factors to the Maspin promoter, both known to activate Maspin transcription. PAR-1 silencing did not affect Ets-1 or c-Jun expression; rather it resulted in increased expression of the chromatin remodeling complex CBP/p300, as well as decreased activity of the CBP/p300 inhibitor p38, resulting in increased binding of Ets-1 and c-Jun to the Maspin promoter and higher Maspin expression. Functionally, Maspin expression reduced the invasive capability of melanoma cells after PAR-1 silencing, which was abrogated after rescuing with PAR-1. Furthermore, tumor growth and experimental lung metastasis was significantly decreased after expressing Maspin in a metastatic melanoma cell line. Moreover, silencing Maspin in PAR-1-silenced cells reverted the inhibition of tumor growth and experimental lung metastasis. Herein, we demonstrate a mechanism by which PAR-1 negatively regulates the expression of the Maspin tumor-suppressor gene in the acquisition of the metastatic melanoma phenotype, thus attributing an alternative function to PAR-1 other than coagulation.
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Prasad CP, Rath G, Mathur S, Bhatnagar D, Ralhan R. Expression analysis of maspin in invasive ductal carcinoma of breast and modulation of its expression by curcumin in breast cancer cell lines. Chem Biol Interact 2009; 183:455-61. [PMID: 19944674 DOI: 10.1016/j.cbi.2009.11.019] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/19/2009] [Revised: 11/19/2009] [Accepted: 11/19/2009] [Indexed: 01/05/2023]
Abstract
In breast cancer, maspin, a serine protease inhibitor, can suppress tumor growth and metastasis in vivo and tumor cell motility and invasion in vitro. The clinical significance of maspin expression in breast cancer, especially in the sequence of ductal carcinoma in situ (DCIS)-invasive cancer-lymph node metastasis is well known in the Western countries, but its status in the rapidly increasing breast cancers in India remains unknown. The present study was designed to determine the clinical significance of maspin expression in invasive ductal carcinomas of breast (IDCs) in North Indian population and modulation of its expression by curcumin. Immunohistochemical analysis of maspin showed loss or reduced cytoplasmic expression in 36 of 59 (61%) tumors. Furthermore, breast cancer cells (MCF-7 (wild type p53) and MDA-MB-231 (mutant p53)) were treated with curcumin and the effect on expression of maspin gene at transcription and translation levels was analyzed by RT-PCR, immunofluorescence and Western blotting. Maspin expression was also correlated with p53 and Bcl-2 levels. Curcumin inhibited cell growth, induced apoptosis and upregulated maspin gene expression in MCF-7 cells and these findings were further correlated with the upregulation of p53 protein and downregulation of Bcl-2, suggesting maspin mediated apoptosis in MCF-7 cells. To our knowledge this is the first report showing the upregulation of maspin expression by curcumin in breast cancer cells and taken together with the clinical data suggests a potential therapeutic role for curcumin in inducing maspin mediated inhibition of invasion of breast carcinoma cells.
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Affiliation(s)
- Chandra P Prasad
- Department of Anatomy, Vardhman Mahavir Medical College & Safdarjang Hospital, New Delhi, India
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Kim SM, Myoung H, Choung PH, Kim MJ, Lee SK, Lee JH. Metastatic leiomyosarcoma in the oral cavity: case report with protein expression profiles. J Craniomaxillofac Surg 2009; 37:454-60. [PMID: 19664933 DOI: 10.1016/j.jcms.2009.06.010] [Citation(s) in RCA: 26] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/17/2009] [Revised: 06/06/2009] [Accepted: 06/11/2009] [Indexed: 10/20/2022] Open
Abstract
Leiomyosarcoma (LMS) is a relatively uncommon malignant tumour derived from smooth muscle cells that rapidly metastasizes to distant regions. It rarely reaches oral tissues in which smooth muscle tissues are absent. We report the case of a 56-year-old woman who presented with LMS in the maxilla that had metastasized from a primary tumour in her uterus, received a total hysterectomy with bilateral salpingo-oophorectomy 9 months earlier. To reveal the poor prognosis of metastatic LMS, a total of 26 antibodies against different factors related to the proliferation, apoptosis, necrosis, and angiogenesis were simultaneously applied on the immunohistochemistry and immuno-blot detection in order to screen for expression n of different proteins in the metastatic LMS. Compared with the immunoreactions of primary uterine LMS, the different antibodies for cellular proliferation, i.e., proliferating cell nuclear antigen (PCNA), multiple primary neoplasm-2 (MPN-2), Max, p21, CDK4, p53, Rb-1, Bad, Bcl-2, epidermal growth factor receptor (EGF-R), hepatocyte growth factor (HGF), C-erbb2, Maspin, and DMBT-1, and those for angiogenesis, i.e., vWF, CD31, and Angiogenin, were more intensely expressed, while Bax, p16, Wnt-1, E-cadherin, and APC were relatively weakly expressed. In particular, beta-catenin was densely localized to the nuclei of tumour cells. These data suggest that rapid proliferation of the tumour cells is related to over-expression of different oncogenes, and that the infiltrative growth and early distant metastasis of these tumour cells are related to over-expression of angiogenesis factors. A total of seven cases of metastatic LMS to the oral cavity that had been published in the English literature were reviewed, and the reason for the poor prognosis in the metastatic LMS is suggested in this case report.
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Affiliation(s)
- Soung Min Kim
- Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, Seoul, Republic of Korea
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36
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Expressions of maspin, P53 and Skp2 in colorectal tumors and their clinicopathological significance. Chin J Cancer Res 2009. [DOI: 10.1007/s11670-009-0147-z] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/20/2022] Open
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Frey A, Soubani AO, Adam AK, Sheng S, Pass HI, Lonardo F. Nuclear, compared with combined nuclear and cytoplasmic expression of maspin, is linked in lung adenocarcinoma to reduced VEGF-A levels and in Stage I, improved survival. Histopathology 2009; 54:590-7. [PMID: 19309490 PMCID: PMC2911575 DOI: 10.1111/j.1365-2559.2009.03260.x] [Citation(s) in RCA: 25] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Abstract
AIMS To evaluate whether there is a correlation between the subcellular localization of maspin and the histological, molecular and biological features of pulmonary adenocarcinoma, particularly addressing the hypothesis that the tumour inhibitor properties of maspin may be linked to a nuclear, compared with a combined nuclear and cytoplasmic expression pattern. METHODS AND RESULTS The subcellular expression of maspin was determined in 80 resected pulmonary adenocarcinomas (Stage I, 46; Stage II, 10; Stage III, 20; Stage IV, 4) and correlated with histological grade, proliferative rate, p53 expression, vascular endothelial growth factor (VEGF)-A levels, and prognosis (mean follow-up of 41.5 months). Cases with nuclear (N) maspin (n = 47), compared with the [N + cytoplasmic (C)] group (n = 28), showed lower (P CONCLUSIONS (N) maspin selects lung adenocarcinomas with distinct molecular and clinical features, supporting the hypothesis that its tumour inhibitor properties may be linked to its nuclear localization.
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Affiliation(s)
- Amy Frey
- Department of Pathology, Harper University Hospital, Wayne State University, Detroit, MI 48201, USA
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38
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Cytoplasmic and nuclear maspin expression in lung carcinomas: an immunohistochemical study using tissue microarrays. Appl Immunohistochem Mol Morphol 2008; 16:459-65. [PMID: 18665036 DOI: 10.1097/pai.0b013e3181640bb1] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
Abstract
Maspin, a serine protease inhibitor related to the serpin family, can inhibit invasion and metastasis of malignancies although direct evidence of the clinicopathologic significance of cytoplasmic relative to nuclear expression is limited. Here, maspin expression was examined on tissue microarrays containing lung carcinoma (n=155) and adjacent noncancerous tissue (n=20) and also 4 lung carcinoma cell lines (LC-1/Sq, LC-IF, PC-14, and AoI) by immunohistochemistry. Maspin expression was compared with clinicopathologic parameters of the tumors. Maspin expression showed positive nuclear staining in basal cells, LC-IF, and PC-14 cell lines, and also cytoplasmic immunoreactivity in secretory and ciliated cells, LC-1/Sq cell line. Cytoplasmic staining was the lowest in adenocarcinoma (AD) and the highest in squamous cell carcinoma as compared with other types of lung carcinoma (P<0.05), and positively correlated with expression of p53 and caspase-3 (P<0.05). The cytoplasmic one showed stronger immunoreactivity in male carcinoma patients than female ones (P<0.05). The nuclear maspin expression gradually increased through squamous cell carcinoma, AD, large cell carcinoma to small cell carcinoma (P<0.05) and was also positively associated with the levels of vascular epithelial growth factor and extracellular matrix metalloproteinase inducer expression (P<0.05). Kaplan-Meier analysis indicated that the cytoplasmic or nuclear maspin expression was not a good prognostic marker for lung carcinomas overall (P>0.05), but the cytoplasmic pattern pointed to good survival for AD cases (P<0.05). It was concluded that the cytoplasmic and nuclear expression patterns of maspin are involved in the cellular differentiation of normal lung tissue and the histogenesis of different lung carcinomas. The cytoplasmic maspin may play an important role in lung carcinomas by regulating apoptosis and thus is a favorable prognostic marker for AD patients, whereas the nuclear location may be linked to promotion of angiogenesis.
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Takanami I, Abiko T, Koizumi S. Expression of Maspin in Non–Small-Cell Lung Cancer: Correlation with Clinical Features. Clin Lung Cancer 2008; 9:361-6. [DOI: 10.3816/clc.2008.n.052] [Citation(s) in RCA: 22] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022]
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Zieglschmid V, Hollmann C, Böcher O. DETECTION OF DISSEMINATED TUMOR CELLS IN PERIPHERAL BLOOD. Crit Rev Clin Lab Sci 2008; 42:155-96. [PMID: 15941083 DOI: 10.1080/10408360590913696] [Citation(s) in RCA: 195] [Impact Index Per Article: 11.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/23/2022]
Abstract
Metastases are the major cause of cancer-related deaths in patients with solid epithelial malignancies, such as breast, colorectal and prostate carcinomas. Hematogenous spreading of tumor cells from a primary tumor can be considered as a crucial step in the metastasis cascade leading eventually to the formation of clinically manifest metastases. Consequently, as shown in recent studies, the detection of disseminated tumor cells in peripheral blood might be of clinical relevance with respect to individual patient prognosis and staging or monitoring of therapy. However, the rarity of disseminated tumor cells in peripheral blood renders the application of sensitive techniques mandatory for their detection. The emergence of highly sophisticated reverse transciptase-polymerase chain reaction (RT-PCR) assays, combining a preanalytical enrichment step with the assessment of multiple molecular tumor markers expressed in disseminated tumor cells, provides a powerful tool in detecting disseminated tumor cells with high sensitivity and specificity. This review will discuss currently used tumor markers as well as experimental means to enhance the sensitivity and specificity of RT-PCR assays to detect disseminated tumor cells in the peripheral blood of patients with breast, colorectal, and prostate cancers, and their clinical relevance assessed in recent studies.
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Horswill MA, Narayan M, Warejcka DJ, Cirillo LA, Twining SS. Epigenetic silencing of maspin expression occurs early in the conversion of keratocytes to fibroblasts. Exp Eye Res 2008; 86:586-600. [PMID: 18291368 PMCID: PMC2374753 DOI: 10.1016/j.exer.2008.01.003] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/15/2007] [Revised: 12/10/2007] [Accepted: 01/03/2008] [Indexed: 10/22/2022]
Abstract
Maspin, a 42 kDa non-classical serpin (serine protease inhibitor) that controls cell migration and invasion, is mainly expressed by epithelial-derived cells but is also expressed in corneal stromal keratocytes. Upon culture of stromal keratocytes in the presence of FBS, maspin is down-regulated to nearly undetectable levels by passage two. DNA methylation is one of several processes that controls gene expression during cell differentiation, development, genetic imprinting, and carcinogenesis but has not been studied in corneal stromal cells. The purpose of this study was to determine whether DNA methylation of the maspin promoter and histone H3 dimethylation is involved in the mechanism of down-regulation of maspin synthesis in human corneal stromal fibroblasts and myofibroblasts. Human donor corneal stroma cells were immediately placed into serum-free defined medium or cultured in the presence of FBS and passed into serum-free medium or medium containing FBS or FGF-2 to induce the fibroblast phenotype or TGF-beta1 for the myofibroblast phenotype. These cell types are found in wounded corneas. The cells were used to prepare RNA for semi-quantitative or quantitative RT-PCR or to extract protein for Western analysis. In addition, P4 FBS cultured fibroblasts were treated with the DNA demethylating agent, 5-aza-2'-deoxycytidine (5-Aza-dC), and the histone deacetylase inhibitor, trichostatin A (TSA). Cells with and without treatment were harvested and assayed for DNA methylation using sodium bisulfite sequencing. The methylation state of histone H3 associated with the maspin gene in the P4 fibroblast cells was determined using a ChIP assay. Freshly harvested corneal stromal cells expressed maspin but upon phenotypic differentiation, maspin mRNA and protein were dramatically down-regulated. Sodium bisulfite sequencing revealed that the maspin promoter in the freshly isolated stromal keratocytes was hypomethylated while both the P0 stromal cells and the P1 cells cultured in the presence of serum-free defined medium, FGF-2 and TGF-beta1 were hypermethylated. Down-regulation of maspin synthesis was also associated with histone H3 dimethylation at lysine 9. Both maspin mRNA and protein were re-expressed at low levels with 5-Aza-dC but not TSA treatment. Addition of TSA to 5-Aza-dC treated cells did not increase maspin expression. Treatment with 5-Aza-dC did not significantly alter demethylation of the maspin promoter but did demethylate histone H3. These results show maspin promoter hypermethylation and histone methylation occur with down-regulation of maspin synthesis in corneal stromal cells and suggest regulation of genes upon conversion of keratocytes to wound healing fibroblasts can involve promoter and histone methylation.
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Affiliation(s)
- Mark A. Horswill
- Department of Biochemistry, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226
| | - Malathi Narayan
- Department of Biochemistry, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226
| | - Debra J. Warejcka
- Department of Biochemistry, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226
| | - Lisa A. Cirillo
- Department of Cellular Biology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226
| | - Sally S. Twining
- Department of Biochemistry, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226
- Department of Ophthalmology, Eye Institute, 925 North 87th Street, Milwaukee, WI 53226
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Bolat F, Gumurdulu D, Erkanli S, Kayaselcuk F, Zeren H, Ali Vardar M, Kuscu E. Maspin overexpression correlates with increased expression of vascular endothelial growth factors A, C, and D in human ovarian carcinoma. Pathol Res Pract 2008; 204:379-87. [PMID: 18343598 DOI: 10.1016/j.prp.2008.01.011] [Citation(s) in RCA: 25] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/14/2007] [Accepted: 01/22/2008] [Indexed: 12/13/2022]
Abstract
The vascular endothelial growth factor (VEGF) family, including VEGFA, VEGFC, and VEGFD, plays an essential role in the angiogenesis of both pathologic and nonpathologic conditions. Maspin belongs to the serpin superfamily and has been identified as a tumor suppressor because it inhibits motility, invasion, and angiogenesis. Few studies have compared maspin with VEGF in ovarian carcinoma. Therefore, we investigated the expression and correlation of maspin, VEGFA, VEGFC, and VEGFD with the tumorigenesis of the ovary and clinicopathologic variables. Using immunohistochemistry, we examined maspin, VEGFA, VEGFC, and VEGFD expression in 60 ovarian carcinoma tissues (35 serous papillary carcinomas, 18 endometrioid carcinomas, and 7 primary ovarian mucinous carcinomas). Staining of cells was scored as +2 if more than 50% of the cells were positive, as +1 if less than 50% of the cells were positive, and as negative if none of the cells stained positive. Overexpression of maspin, VEGFC, and VEGFD was significantly associated with high tumor grade (P<.001, P=.004, P<.001, respectively), clinical stage (P=.002, .01, and .001, respectively), the presence of ascites (P<.001, P=.03, and P=.001, respectively), and the presence of metastatic lymph nodes (P=.002, P<.001, and P<.001, respectively). Maspin was correlated with VEGFA (P=.01), VEGFC (P<.001), and VEGFD (P<.001). The VEGFA score was positively correlated with high tumor grade (P=.04), lymphovascular space invasion (LVSI) (P<.001), International Federation of Gynecology and Obstetrics (FIGO) stage (P=.009), maspin, VEGFC (P=.003), and VEGFD (P=.003), but it was not correlated with the presence of ascites and metastatic lymph nodes. Maspin, VEGFC, and VEGFD are expressed in ovarian tumors with a poor prognostic parameters, and seem to play a role in ovarian cancer angiogenesis, progression, and lymph node metastases. Our results indicate that in contrast to most other carcinomas, maspin expression is directly associated with the biological aggressiveness of ovarian carcinoma. These results may offer new insights regarding the role of maspin in ovarian cancer and might also affect the diagnosis and treatment strategies.
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Affiliation(s)
- Filiz Bolat
- Baskent University Faculty of Medicine, Department of Pathology, Ankara, Turkey.
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Yu M, Zheng H, Tsuneyama K, Takahashi H, Nomoto K, Xu H, Takano Y. Paradoxical expression of maspin in gastric carcinomas: correlation with carcinogenesis and progression. Hum Pathol 2007; 38:1248-55. [PMID: 17490717 DOI: 10.1016/j.humpath.2006.11.025] [Citation(s) in RCA: 21] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/19/2006] [Revised: 11/22/2006] [Accepted: 11/28/2006] [Indexed: 11/21/2022]
Abstract
Maspin, a serine protease inhibitor related to the serpin family, can suppress invasion and metastasis of malignancies. To clarify the role of maspin in the genesis and progression of gastric carcinomas, its expression pattern and level were studied by immunohistochemistry on tissue microarrays containing gastric carcinoma (n = 237), normal gastric mucosa (n = 23), intestinal metaplasia (n = 38), and adenoma (n = 42); and the findings were compared with clinicopathological parameters. Furthermore, maspin expression in the gastric carcinoma cell lines (HCG-27, MKN28, and MKN45) was examined by immunohistochemistry and Western blotting. We found that cytoplasmic and nuclear maspin expression paralleled each other (P < .05) and decreased from intestinal metaplasia, adenoma, and carcinoma to normal gastric mucosa (P < .05). A significant positive association was noted with depth of invasion, lymphatic invasion, lymph node metastasis, and TNM stage (P < .05) but not with sex or Lauren's classification (P > .05). Univariate and multivariate analyses indicated that expression of maspin correlated negatively with cumulative patient survival in gastric carcinoma (P < .05) but was not an independent factor in the prognosis. The 2 independent factors, depth of invasion and lymphatic invasion, influenced the relation between nuclear maspin expression and survival, whereas only depth of invasion correlated with cytoplasmic maspin. Our study indicated that maspin expression experiences upregulation in gastric precancerous lesions and then slight downregulation with malignant transformation. High expression may paradoxically promote invasion and metastasis of gastric carcinomas and could be considered a good marker for the pathobiological behaviors of gastric carcinomas.
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Affiliation(s)
- Miao Yu
- Department of Diagnostic Pathology, Graduate School of Medicine and Pharmaceutical Science, University of Toyama, Sugitani, Toyama 2630, Japan
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Denk AE, Bettstetter M, Wild PJ, Hoek K, Bataille F, Dietmaier W, Bosserhoff AK. Loss of maspin expression contributes to a more invasive potential in malignant melanoma. ACTA ACUST UNITED AC 2007; 20:112-9. [PMID: 17371437 DOI: 10.1111/j.1600-0749.2007.00363.x] [Citation(s) in RCA: 34] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/01/2022]
Abstract
Deregulation of protease expression and activity is known to play an important role in tumour progression of malignant melanoma. The serpin maspin, a tumour suppressor in breast and prostate cancer was described as an inhibitor of cell migration and inducer of cell adhesion between the basement membrane and extracellular matrix resulting in inhibition of tumour metastasis. In contrast, overexpression of maspin is correlated with poor prognosis in other cancers. However, little is known about expression, regulation and function of maspin in malignant melanoma. In this study, we found loss of maspin expression in malignant melanoma cells compared with normal human epidermal melanocytes, which was analysed by quantitative real-time PCR, Western blot analysis, immunohistochemistry and microarray. For functional studies, melanoma cell clones stably transfected with a maspin expression vector were tested for changes in proliferation, migration and invasion. Although we could not see differences in proliferation and migration, we detected strongly reduced invasive capacity in the melanoma cell clones in which maspin is re-expressed compared with control. Reduced invasive potential was also detected in three different melanoma cell lines transiently transfected with a maspin expression vector. Furthermore, exogenously added maspin alone was sufficient to reduce invasion in MelIm significantly, indicating that maspin directly inhibits invasion on the cell surface. In summary, we believe that maspin is a tumour suppressor in malignant melanoma.
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Affiliation(s)
- Alexandra E Denk
- Institute of Pathology, University of Regensburg, Regensburg, Germany
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Cui XN, Tang JW, Hou L, Song B, Ban LY. Identification of differentially expressed genes in mouse hepatocarcinoma ascites cell line with low potential of lymphogenous metastasis. World J Gastroenterol 2006; 12:6893-7. [PMID: 17106944 PMCID: PMC4087450 DOI: 10.3748/wjg.v12.i42.6893] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/07/2023] Open
Abstract
AIM: To identify genes differentially expressed in mouse hepatocarcinoma ascites cell line with low potential of lymphogenous metastasis.
METHODS: A subtracted cDNA library of mouse hepatocarcinoma cell line with low potential of lympho-genous metastasis Hca-P and its synogenetic cell line Hca-F with high metastatic potential was constructed by suppression subtracted hybridization (SSH) method. The screened clones of the subtracted library were sequenced and GenBank homology search was performed.
RESULTS: Fifteen differentially expressed cDNA fragments of Hca-P were obtained which revealed 8 known genes, 4 expressed sequence tags (ESTs) and 3 cDNAs showed no homology.
CONCLUSION: Tumor metastasis is an incident involving multiple genes. SSH is a useful technique to detect differentially expressed genes and an effective method to clone novel genes.
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MESH Headings
- Animals
- Ascites/pathology
- Carcinoma, Hepatocellular/genetics
- Carcinoma, Hepatocellular/pathology
- Cell Line, Tumor
- Deoxyribonucleases, Type II Site-Specific/metabolism
- Gene Expression Profiling
- Gene Expression Regulation, Neoplastic
- Genes, Tumor Suppressor
- Hybridization, Genetic
- Liver Neoplasms/genetics
- Liver Neoplasms/pathology
- Lymphatic Metastasis/genetics
- Mice
- Mice, Inbred Strains
- RNA, Messenger/genetics
- Suppression, Genetic
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Affiliation(s)
- Xiao-Nan Cui
- Department of Oncology, The 1st Affiliated Dalian Medical University, Dalian 116027, Liaoning Province, China
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Abstract
Maspin, a mammary serine protease inhibitor, was originally reported as a tumor suppressor gene in breast cancer. The purpose of the present study was to examine maspin expression and evaluate its clinicopathological significance in endometrial cancer. We examined maspin expression immunohistochemically in 41 cases with endometrioid adenocarcinoma. DNA methylation status at the maspin promoter region was determined by the methylation-specific polymerase chain reaction method. Aberrant maspin expression was observed in 27 (66%) of 41 endometrioid adenocarcinomas but not in normal endometrial glands. Maspin immunoreactivity of the tumor cells varied in incidence and density among tumors. Positive staining was correlated significantly with the presence of squamous differentiation (presence vs absence = 11/11 [100%] vs 16/30 [53%], P < 0.05), and nuclear subcellular localization of maspin protein was also significantly associated with squamous differentiation (nuclear positive vs nuclear negative = 6/11 [54%] vs 2/30 [6.7%], P < 0.05). An inverse correlation between their immunoreactivity and methylation status was observed (P < 0.01). Three of the four cell lines established from endometrioid adenocarcinomas overexpressed maspin mRNA and its protein product. In a maspin-negative cell line, maspin expression was induced by treatment with 5-aza-2'-deoxycytidine, a DNA demethylating agent. There was no significant correlation between maspin expression and any clinicopathlogical data. These findings suggest that maspin induced by DNA demethylation at the promoter region may contribute to squamous differentiation of tumor cells in endometrioid adenocarcinomas.
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Affiliation(s)
- Shinya Murai
- Department of Obstetrics and Gynecology, School of Medicine, Iwate Medical University, Uchimaru 19-1, Morioka, Iwate 020-8505, Japan
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Secord AA, Lee PS, Darcy KM, Havrilesky LJ, Grace LA, Marks JR, Berchuck A. Maspin expression in epithelial ovarian cancer and associations with poor prognosis: A Gynecologic Oncology Group study. Gynecol Oncol 2006; 101:390-7. [PMID: 16551475 DOI: 10.1016/j.ygyno.2006.02.014] [Citation(s) in RCA: 25] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/29/2005] [Revised: 02/07/2006] [Accepted: 02/08/2006] [Indexed: 01/06/2023]
Abstract
OBJECTIVE This study examined MASPIN expression in human ovarian cancer, and explored the association between MASPIN and prognosis in patients with advanced stage disease treated with first-line cisplatin, carboplatin and/or paclitaxel. METHODS Frozen primary tumors were obtained from 68 women with previously untreated, advanced stage epithelial ovarian cancer who participated in a specimen banking protocol and a phase III treatment trial conducted by the Gynecologic Oncology Group. Immunoblot analysis was performed in lysates prepared from these tumor specimens to quantify the relative expression of MASPIN/beta-actin. RESULTS MASPIN was expressed at detected levels in 49 (72%) cases with relative expression ranging from 0.02 to 7.7 (median = 0.2), and was not detected in 19 (28%) of the primary tumors tested. Non-detectable levels of this class II tumor suppressor gene product and inhibitor of angiogenesis were associated with suboptimally-debulked disease (P = 0.034) but not with patient age, FIGO stage, tumor grade, or histologic subtype. After adjusting for prognostic variables for disease progression or death, non-detectable MASPIN expression predicted an increased risk of disease progression (hazard ratio [HR] = 1.89; 95% confidence interval [CI]: 1.04-3.45; P = 0.038) and death (HR = 1.99; 95% CI: 1.07-3.69; P = 0.030). CONCLUSIONS In advanced stage epithelial ovarian cancer, non-detectable MASPIN appears to be associated with suboptimally-debulked disease and be an independent predictor of an increased risk of progression and death. Further studies are needed to validate these exploratory findings, determine the molecular mechanism controlling MASPIN expression as well as down-regulation and loss in ovarian cancer, and determine if MASPIN can prevent progression of this disease.
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Dietmaier W, Bettstetter M, Wild PJ, Woenckhaus M, Rümmele P, Hartmann A, Dechant S, Blaszyk H, Pauer A, Klinkhammer-Schalke M, Hofstädter F. Nuclear Maspin expression is associated with response to adjuvant 5-fluorouracil based chemotherapy in patients with stage III colon cancer. Int J Cancer 2006; 118:2247-54. [PMID: 16331619 DOI: 10.1002/ijc.21620] [Citation(s) in RCA: 38] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
Maspin, a member of the Serpin protease inhibitor family, is overexpressed in poorly differentiated colorectal tumors and more frequently found in tumors with microsatellite instability. Immunohistochemical nuclear Maspin staining is predominantly seen in tumor cells at the invasion front of such cancers, suggesting that this molecule is associated with local tumor cell infiltration and aggressiveness. In a retrospective study, we studied nuclear Maspin expression as a potential prognostic tool in a total of 172 primary stage III colon cancers by immunohistochemistry. Of those 172 patients, 76 were treated by surgery only, and 96 patients received additional adjuvant 5-fluorouracil (5-FU) based chemotherapy. Nuclear Maspin expression was an independent adverse prognostic factor for overall survival in our patient cohort (hazard ratio 2.08; 95% CI, 1.13-3.81; p = 0.018). However, patients with primary tumors expressing Maspin in the nucleus showed a significant treatment benefit from 5-FU chemotherapy (hazard ratio 0.384; 95% CI, 0.188-0.784; p = 0.009) compared to adjuvantly treated patients whose tumors did not express this molecule. Nuclear Maspin expression is highly predictive of 5-FU chemotherapy response in patients with advanced stage colon cancer. Patients with negative immunohistochemical Maspin expression do not benefit from 5-FU treatment and may be candidates for an alternative (non-5-FU based) adjuvant therapy regime.
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Affiliation(s)
- Wolfgang Dietmaier
- Department of Pathology and Molecular Diagnostics, University of Regensburg, Germany.
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Li HWR, Leung SW, Cheung ANY, Yu MMY, Chan LKY, Wong YF. Expression of maspin in gestational trophoblastic disease. Gynecol Oncol 2006; 101:76-81. [PMID: 16271752 DOI: 10.1016/j.ygyno.2005.09.037] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/12/2005] [Revised: 09/14/2005] [Accepted: 09/22/2005] [Indexed: 10/25/2022]
Abstract
BACKGROUND Maspin is a tumor suppressor gene whose expression is altered in neoplasia and malignancies of many tissues. In the human placenta, the maspin gene is expressed in trophoblastic cells and might act as an inhibitory regulator of trophoblastic invasion. Hence, in gestational trophoblastic disease (GTD), where there is increased propensity for invasion in the trophoblastic tissue, we hypothesized that maspin expression would be decreased. The present study aimed at investigating the expression of maspin in GTD and its prognostic significance. METHODS Using immunohistochemical staining, we firstly studied the expression of maspin in hydatidiform moles, with gestational age-matched normal first trimester placenta used as control. A total of 38 cases of hydatidiform moles were studied, including 20 complete moles (CM) and 18 partial moles (PM). Among them, 10 cases of the CM group and 8 cases of the PM group subsequently developed gestational trophoblastic neoplasia (GTN). Immunostaining was also performed on tissue from 4 cases of choriocarcinoma and 5 cases of placental site trophoblastic tumor. Reverse transcriptase-polymerase chain reaction (RT-PCR) was further performed on RNA extracted from 10 hydatidiform moles (5 with GTN and 5 without) and 6 normal first-trimester placentae. RESULTS In all tissue sections, nuclear expression of immunostaining signal was demonstrated, mainly in the cytotrophoblasts. The percentage of trophoblastic nuclei stained in both complete and partial moles was significantly lower than that in normal first-trimester placenta (P < 0.001). However, there was no significant difference in immunostaining between complete and partial moles (P > 0.05). There was also significantly lower expression of maspin in those cases subsequently developing GTN than those which did not (P = 0.01). Immunostaining on choriocarcinoma and placental site trophoblastic tumor showed reduced expression of maspin in all the tumor cells. Reverse transcriptase-polymerase chain reaction revealed that the expression of maspin was consistently down-regulated in all the hydatidiform mole samples. CONCLUSIONS Our results suggest that there is down-regulated expression of maspin in gestational trophoblastic diseases, and the down-regulation is more prominent in cases developing gestational trophoblastic neoplasia. This may play a role with prognostic significance in the pathogenesis and malignant transformation of hydatidiform moles.
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Affiliation(s)
- H W Raymond Li
- Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, New Territories, Hong Kong.
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Abstract
Maspin is a unique serine proteinase inhibitor that has tumor suppressor activity. It has been reported that maspin is expressed in normal human mammary epithelial cells and it is down-regulated during the progression of cancer. However, to date, there is very limited data on the clinical significance of maspin expression in human breast cancer. In this study, maspin expression was assessed immunohistochemically from 80 invasive ductal carcinoma (IDC) specimens of the breast. Also, maspin expression was compared with the clinicopathological factors (age, grade, tumor size and lymph node status), the expression of estrogen receptor (ER), progesterone receptor (PR) and p53, DNA ploidy and the overall survival in an attempt to assess its prognostic value. The maspin expression was positive in 25 IDC cases (31.3%). The maspin expression in IDC was significantly correlated with a higher histologic grade, a larger tumor size, a positive p53 status and shorter survival. There was an inverse association with maspin expression and the PR status. These findings suggest that maspin expression is not down-regulated with the progression of cancer and maspin expression may be associated with a poor prognosis. The immunohistochemical detection of maspin in breast cancers may be helpful for predicting an aggressive phenotype.
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MESH Headings
- Adult
- Aged
- Breast Neoplasms/genetics
- Breast Neoplasms/metabolism
- Breast Neoplasms/mortality
- Breast Neoplasms/pathology
- Carcinoma, Ductal, Breast/genetics
- Carcinoma, Ductal, Breast/metabolism
- Carcinoma, Ductal, Breast/mortality
- Carcinoma, Ductal, Breast/pathology
- DNA, Neoplasm/analysis
- DNA, Neoplasm/genetics
- Female
- Genes, Tumor Suppressor
- Humans
- Middle Aged
- Ploidies
- Prognosis
- Receptors, Estrogen/metabolism
- Receptors, Progesterone/metabolism
- Serpins/metabolism
- Survival Rate
- Tumor Suppressor Protein p53/metabolism
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Affiliation(s)
- Mi Ja Lee
- Department of Pathology, College of Medicine, Chosun University, Dong-gu, Gwangju, Korea.
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