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Vakhrusheva O, Zhao F, Markowitsch SD, Slade KS, Brandt MP, Tsaur I, Cinatl J, Michaelis M, Efferth T, Blaheta RA, Haferkamp A, Juengel E. Artesunate Inhibits Metastatic Potential in Cisplatin-Resistant Bladder Cancer Cells by Altering Integrins. Cells 2025; 14:570. [PMID: 40277897 PMCID: PMC12026051 DOI: 10.3390/cells14080570] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/20/2025] [Revised: 03/28/2025] [Accepted: 04/08/2025] [Indexed: 04/26/2025] Open
Abstract
The survival of patients with locally advanced and metastatic bladder cancer (BCa) is persistently low. Hence, new treatment options are urgently needed. Artesunate (ART) a derivative of artemisinin, used in Traditional Chinese Medicine, shows anti-tumor activity extending over a broad spectrum of human cancers. As we have previously shown, ART inhibits growth in cisplatin-sensitive (parental) and cisplatin-resistant BCa cells. However, how ART acts on the metastatic potential of BCa remained unclear. To clarify, we applied ART to parental and cisplatin-resistant RT4, RT112, T24, and TCCSup BCa cell lines. We examined tumor cell adhesion to vascular endothelium and immobilized collagen and evaluated chemotactic activity, migration, and invasive activity of the BCa cells. Adhesion receptors, integrin α and β subtypes, integrin-linked kinase (ILK), and focal adhesion kinase (FAK) were investigated. The functional relevance of integrin expression altered by ART was determined by blocking studies. ART significantly reduced tumor cell adhesion to vascular endothelium and immobilized collagen in parental as well as in cisplatin-resistant BCa cells. Depending on cell type, ART suppressed tumor cell motility and diminished integrin expression (surface and total). Functional blocking of integrins altered by ART reduced cell adhesion and invasion of the BCa cells. Thus, the metastatic potential of parental and cisplatin-resistant BCa cells was significantly inhibited by ART, making it a promising treatment option for patients with advanced or therapy-resistant BCa.
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Affiliation(s)
- Olesya Vakhrusheva
- Department of Urology and Pediatric Urology, University Medical Center Mainz, Langenbeckstr. 1, 55131 Mainz, Germany; (O.V.); (F.Z.); (S.D.M.); (K.S.S.); (M.P.B.); (I.T.); (R.A.B.); (A.H.)
- Department of Urology, University Hospital Tübingen, Hoppe-Seyler-Strasse 3, 72076 Tübingen, Germany
| | - Fuguang Zhao
- Department of Urology and Pediatric Urology, University Medical Center Mainz, Langenbeckstr. 1, 55131 Mainz, Germany; (O.V.); (F.Z.); (S.D.M.); (K.S.S.); (M.P.B.); (I.T.); (R.A.B.); (A.H.)
| | - Sascha Dennis Markowitsch
- Department of Urology and Pediatric Urology, University Medical Center Mainz, Langenbeckstr. 1, 55131 Mainz, Germany; (O.V.); (F.Z.); (S.D.M.); (K.S.S.); (M.P.B.); (I.T.); (R.A.B.); (A.H.)
| | - Kimberly Sue Slade
- Department of Urology and Pediatric Urology, University Medical Center Mainz, Langenbeckstr. 1, 55131 Mainz, Germany; (O.V.); (F.Z.); (S.D.M.); (K.S.S.); (M.P.B.); (I.T.); (R.A.B.); (A.H.)
| | - Maximilian Peter Brandt
- Department of Urology and Pediatric Urology, University Medical Center Mainz, Langenbeckstr. 1, 55131 Mainz, Germany; (O.V.); (F.Z.); (S.D.M.); (K.S.S.); (M.P.B.); (I.T.); (R.A.B.); (A.H.)
| | - Igor Tsaur
- Department of Urology and Pediatric Urology, University Medical Center Mainz, Langenbeckstr. 1, 55131 Mainz, Germany; (O.V.); (F.Z.); (S.D.M.); (K.S.S.); (M.P.B.); (I.T.); (R.A.B.); (A.H.)
- Department of Urology, University Hospital Tübingen, Hoppe-Seyler-Strasse 3, 72076 Tübingen, Germany
| | - Jindrich Cinatl
- Interdisciplinary Laboratory for Paediatric Tumour and Virus Research, Dr. Petra Joh Research Institute, 60529 Frankfurt am Main, Germany; (J.C.J.); (M.M.)
| | - Martin Michaelis
- Interdisciplinary Laboratory for Paediatric Tumour and Virus Research, Dr. Petra Joh Research Institute, 60529 Frankfurt am Main, Germany; (J.C.J.); (M.M.)
- School of Natural Sciences, University of Kent, Canterbury CT2 7NJ, UK
| | - Thomas Efferth
- Institute of Pharmaceutical and Biomedical Sciences, Johannes Gutenberg University Mainz, Staudinger Weg 5, 55128 Mainz, Germany;
| | - Roman Alexander Blaheta
- Department of Urology and Pediatric Urology, University Medical Center Mainz, Langenbeckstr. 1, 55131 Mainz, Germany; (O.V.); (F.Z.); (S.D.M.); (K.S.S.); (M.P.B.); (I.T.); (R.A.B.); (A.H.)
| | - Axel Haferkamp
- Department of Urology and Pediatric Urology, University Medical Center Mainz, Langenbeckstr. 1, 55131 Mainz, Germany; (O.V.); (F.Z.); (S.D.M.); (K.S.S.); (M.P.B.); (I.T.); (R.A.B.); (A.H.)
| | - Eva Juengel
- Department of Urology and Pediatric Urology, University Medical Center Mainz, Langenbeckstr. 1, 55131 Mainz, Germany; (O.V.); (F.Z.); (S.D.M.); (K.S.S.); (M.P.B.); (I.T.); (R.A.B.); (A.H.)
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Immune Exclusion Is Frequent in Small-Cell Carcinoma of the Bladder. DISEASE MARKERS 2019; 2019:2532518. [PMID: 31191745 PMCID: PMC6525886 DOI: 10.1155/2019/2532518] [Citation(s) in RCA: 14] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 02/06/2019] [Accepted: 03/07/2019] [Indexed: 01/03/2023]
Abstract
Small-cell cancer of the urinary bladder is a rare but highly aggressive disease. It is currently unclear whether immune checkpoint therapies that have been approved for urothelial carcinomas will also be efficient in small-cell carcinomas. In this study, we analyzed potential predictors of response including PD-L1 expression and the quantity and location of tumor-infiltrating lymphocytes (TILs) in 12 small-cell and 69 “classical” urothelial cancers by immunohistochemistry. The analysis revealed that small-cell carcinomas were characterized by the virtual absence of PD-L1 expression and an “immune-excluded” phenotype with only a few TILs in the center of the tumor (CT). In small-cell carcinomas, the average immune cell density in the CT (CD3: 159 ± 206, CD8: 87 ± 169 cells/mm2) was more than 3 times lower than that in the urothelial carcinomas (CD3: 625 ± 800, p < 0.001; CD8: 362 ± 626 cells/mm2, p = 0.004) while there was no significant difference in the immune cell density at the invasive margin (IM) (small-cell carcinomas CD3: 899 ± 733, CD8: 404 ± 433 cells/mm2; urothelial carcinomas CD3: 1167 ± 1206, p = 0.31; CD8: 582 ± 864 cells/mm2, p = 0.27). Positive PD-L1 staining was found in 39% of urothelial cancers, but in only 8% of small-cell bladder cancer cases (p = 0.04). Concordant with these data, a sharp decrease of PD-L1 positivity from >80% to 0% positive cells and of TILS in the CT from 466-1063 CD3-positive cells/mm2 to 50-109 CD3-positive cells/mm2 was observed in two cancers with clear-cut progression from “classical” urothelial to small-cell carcinoma. In conclusion, these data demonstrate that small-cell bladder cancer commonly exhibits an immune-excluded phenotype.
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Jennek S, Mittag S, Reiche J, Westphal JK, Seelk S, Dörfel MJ, Pfirrmann T, Friedrich K, Schütz A, Heinemann U, Huber O. Tricellulin is a target of the ubiquitin ligase Itch. Ann N Y Acad Sci 2017; 1397:157-168. [DOI: 10.1111/nyas.13349] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/30/2017] [Revised: 03/08/2017] [Accepted: 03/14/2017] [Indexed: 01/05/2023]
Affiliation(s)
- Susanne Jennek
- Department of Biochemistry II; Jena University Hospital, Friedrich Schiller University Jena; Jena Germany
| | - Sonnhild Mittag
- Department of Biochemistry II; Jena University Hospital, Friedrich Schiller University Jena; Jena Germany
| | - Juliane Reiche
- Department of Biochemistry II; Jena University Hospital, Friedrich Schiller University Jena; Jena Germany
| | - Julie K. Westphal
- Department of Biochemistry II; Jena University Hospital, Friedrich Schiller University Jena; Jena Germany
| | - Stefanie Seelk
- Department of Biochemistry II; Jena University Hospital, Friedrich Schiller University Jena; Jena Germany
| | - Max J. Dörfel
- Department of Biochemistry II; Jena University Hospital, Friedrich Schiller University Jena; Jena Germany
| | - Thorsten Pfirrmann
- Institute of Physiological Chemistry, University Hospital Halle; Martin Luther University Halle-Wittenberg; Halle/Saale Germany
| | - Karlheinz Friedrich
- Department of Biochemistry II; Jena University Hospital, Friedrich Schiller University Jena; Jena Germany
| | - Anja Schütz
- Helmholtz Protein Sample Production Facility; Max-Delbrück-Center for Molecular Medicine; Berlin Germany
| | - Udo Heinemann
- Helmholtz Protein Sample Production Facility; Max-Delbrück-Center for Molecular Medicine; Berlin Germany
- Crystallography; Max Delbrück Center for Molecular Medicine; Berlin Germany
- Chemistry and Biochemistry Institute; Freie Universität Berlin; Berlin Germany
| | - Otmar Huber
- Department of Biochemistry II; Jena University Hospital, Friedrich Schiller University Jena; Jena Germany
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Kouzegaran S, Shahraki K, Makateb A, Shahri F, Hatami N, Behnod V, Tanha AS. Prognostic Investigations of Expression Level of Two Genes FasL and Ki-67 as Independent Prognostic Markers of Human Retinoblastoma. Oncol Res 2016; 25:471-478. [PMID: 27625332 PMCID: PMC7841034 DOI: 10.3727/096504016x14721217330657] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/17/2023] Open
Abstract
In this study, expression of FasL and Ki-67 messenger RNA (FasL and Ki-67 mRNA) in human retinoblastoma (HRB) was examined by the immunohistochemistry method and quantitative real-time PCR. Positive expression of Ki-67 in tumor cells was detected in 16 of 30 patients (53.33%), and only 9 (30%) of the tissues from patients with retinoblastoma showed positive staining for FasL. Our results revealed that FasL expression was significantly higher in tumor tissue with invasion compared with the noninvasion form (p = 0.033). Ki-67 expression was markedly increased in tumor tissues with invasion compared with the noninvasion group (p = 0.04), but no significant correlation was found between FasL expression and differentiation (p > 0.05). In addition, Ki-67 expression was strongly linked to differentiation (p < 0.002). Expression of these FasL was correlated with shorter overall survival of patients, but its expression was not significantly associated with overall survival (p = 0.15). The impact of Ki-67 expression on survival in patients was also evaluated. Ki-67 expression level was not found to be significantly associated with shorter survival (Kaplan–Meier; p = 0.09). Univariate analysis revealed that massive choroidal invasion was correlated with poor prognosis. Taken together, the data suggest that massive choroidal invasion is also an important indicator of poor prognosis for HRB.
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Affiliation(s)
- Samaneh Kouzegaran
- Department of Pediatrics, Faculty of Medicine, Birjand University of Medical Sciences, Birjand, Iran
| | - Kourosh Shahraki
- Department of Ophthalmology, Alzahra Eye Hospital, Zahedan University of Medical Sciences, Zahedan, Iran
| | - Ali Makateb
- Department of Ophthalmology, AJA University of Medical Sciences, Tehran, Iran
| | - Farkhondeh Shahri
- Department of Optometry, School of Rehabilitation, Zahedan University of Medical Sciences, Zahedan, Iran
| | - Negin Hatami
- Department of Radiology, Stanford University, School of Medicine, Stanford, CA, USA
| | - Vahid Behnod
- Department of Molecular Biology, Baqiyatallah University of Medical Sciences, Tehran, Iran
| | - Amir Saber Tanha
- Department of Anesthesia, Faculty of Medicine, Birjand University of Medical Sciences, Birjand, Iran
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Grimm S, Jennek S, Singh R, Enkelmann A, Junker K, Rippaus N, Berndt A, Friedrich K. Malignancy of bladder cancer cells is enhanced by tumor-associated fibroblasts through a multifaceted cytokine-chemokine loop. Exp Cell Res 2015; 335:1-11. [PMID: 25911129 DOI: 10.1016/j.yexcr.2015.04.001] [Citation(s) in RCA: 21] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/11/2014] [Revised: 11/04/2014] [Accepted: 11/11/2014] [Indexed: 12/11/2022]
Abstract
The microenvironment of tumor cells is critically involved in tumor development and progression. Tumor-associated fibroblasts (TAFs) represent a major constituent of the tumor stroma. Tumor cells are operative in the activation of TAFs, whereas TAFs in turn contribute to tumor cell malignancy. This report describes mechanisms of communication between fibroblasts and urinary bladder cancer (UBC) cells. Migration of bladder cancer cell lines RT112 and Cal-29, representing two different grades of dedifferentiation, was enhanced by cocultivation with TAFs. Conditioned medium from tumor cells induced the release of interleukin (IL)-8, hepatocyte growth factor (HGF), matrix metalloproteinase-2, granulocyte macrophage colony-stimulating factor, and monocyte chemotactic protein (MCP)-1 by TAFs. Tumor cell-derived IL-1α was identified as a major mediator of these stimulatory effects. Fibroblasts, on the other hand, exerted a migration and invasion stimulating influence on UBC cells. MCP-1 and HGF were shown to promote cell migration of both bladder cancer cell lines.
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Affiliation(s)
- Susanne Grimm
- Jena University Hospital, Institute of Biochemistry II, Jena, Germany
| | - Susanne Jennek
- Jena University Hospital, Institute of Biochemistry II, Jena, Germany
| | - Rajan Singh
- Jena University Hospital, Institute of Biochemistry II, Jena, Germany
| | | | - Kerstin Junker
- Jena University Hospital, Department of Urology, Jena, Germany
| | - Nora Rippaus
- Jena University Hospital, Institute of Biochemistry II, Jena, Germany
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Thompson DB, Siref LE, Feloney MP, Hauke RJ, Agrawal DK. Immunological basis in the pathogenesis and treatment of bladder cancer. Expert Rev Clin Immunol 2015; 11:265-79. [PMID: 25391391 PMCID: PMC4637163 DOI: 10.1586/1744666x.2015.983082] [Citation(s) in RCA: 44] [Impact Index Per Article: 4.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/02/2023]
Abstract
The pathogenesis and transition of normal urothelium into bladder carcinoma are multifactorial processes. Chronic inflammation causes initiation and progression of the underlying pathophysiology of invasive and metastatic cancer. A dichotomy is observed in the role of immune cells in bladder cancer. While the immune response defends the host by suppressing neoplastic growth, several immune cells, including neutrophils, macrophages and T-lymphocytes, promote tumor development and progression. The levels of human neutrophil peptide-1, -2 and -3, produced by neutrophils, increase in bladder cancer and might promote tumor angiogenesis and growth. The effect of macrophages is primarily mediated by pro-inflammatory cytokines, IL-6 and TNF-α. In addition, the underlying immunological mechanisms of two treatments, BCG and cytokine gene-modified tumor vaccines, and future directions are critically discussed.
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Affiliation(s)
- David B Thompson
- Center for Clinical and Translational Science, Creighton University School of Medicine, CRISS II Room 510, 2500 California Plaza, Omaha, NE 68178, USA
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Knievel J, Schulz WA, Greife A, Hader C, Lübke T, Schmitz I, Albers P, Niegisch G. Multiple mechanisms mediate resistance to sorafenib in urothelial cancer. Int J Mol Sci 2014; 15:20500-17. [PMID: 25387078 PMCID: PMC4264180 DOI: 10.3390/ijms151120500] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/25/2014] [Revised: 10/30/2014] [Accepted: 10/30/2014] [Indexed: 12/24/2022] Open
Abstract
Genetic and epigenetic changes in the mitogen activated protein kinase (MAPK) signaling render urothelial cancer a potential target for tyrosine kinase inhibitor (TKI) treatment. However, clinical trials of several TKIs failed to prove efficacy. In this context, we investigated changes in MAPK signaling activity, downstream apoptotic regulators and changes in cell cycle distribution in different urothelial cancer cell lines (UCCs) upon treatment with the multikinase inhibitor sorafenib. None of the classical sorafenib targets (vascular endothelial growth factor receptor 1/-receptor 2, VEGFR1/-R2; platelet-derived growth factor receptor α/-receptor β, PDGFR-α/-β; c-KIT) was expressed at significant levels leaving RAF proteins as its likely molecular target. Low sorafenib concentrations paradoxically increased cell viability, whereas higher concentrations induced G1 arrest and eventually apoptosis. MAPK signaling remained partly active after sorafenib treatment, especially in T24 cells with an oncogenic HRAS mutation. AKT phosphorylation was increased, suggesting compensatory activation of the phosphatidylinositol-3-kinase (PI3K) pathway. Sorafenib regularly down regulated the anti-apoptotic myeloid cell leukemia 1 (Mcl-1) protein, but combinatorial treatment with ABT-737 targeting other B-cell lymphoma 2 (Bcl-2) family proteins did not result in synergistic effects. In summary, efficacy of sorafenib in urothelial cancer cell lines appears hampered by limited effects on MAPK signaling, crosstalk with further cancer pathways and an anti-apoptotic state of UCCs. These observations may account for the lack of efficacy of sorafenib in clinical trials and should be considered more broadly in the development of signaling pathway inhibitors for drug therapy in urothelial carcinoma.
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Affiliation(s)
- Judith Knievel
- Department of Urology, Heinrich-Heine-University, Moorenstr. 5, Düsseldorf D-40225, Germany.
| | - Wolfgang A Schulz
- Department of Urology, Heinrich-Heine-University, Moorenstr. 5, Düsseldorf D-40225, Germany.
| | - Annemarie Greife
- Department of Urology, Heinrich-Heine-University, Moorenstr. 5, Düsseldorf D-40225, Germany.
| | - Christiane Hader
- Department of Urology, Heinrich-Heine-University, Moorenstr. 5, Düsseldorf D-40225, Germany.
| | - Tobias Lübke
- Helmholtz-Zentrum für Infektionsforschung, Inhoffenstr. 7, Braunschweig D-38124, Germany.
| | - Ingo Schmitz
- Helmholtz-Zentrum für Infektionsforschung, Inhoffenstr. 7, Braunschweig D-38124, Germany.
| | - Peter Albers
- Department of Urology, Heinrich-Heine-University, Moorenstr. 5, Düsseldorf D-40225, Germany.
| | - Günter Niegisch
- Department of Urology, Heinrich-Heine-University, Moorenstr. 5, Düsseldorf D-40225, Germany.
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Bartsch G, Mitra AP, Cote RJ. Expression profiling for bladder cancer: strategies to uncover prognostic factors. Expert Rev Anticancer Ther 2011; 10:1945-54. [PMID: 21110760 DOI: 10.1586/era.10.131] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/01/2023]
Abstract
Despite being a common cancer worldwide, management of transitional cell carcinoma of the bladder currently relies primarily on clinical staging and histopathologic parameters. Assaying alterations in molecular pathways can contribute valuable information that can accurately predict outcome and chemotherapeutic response in individual patients with bladder cancer. Medium- to high-throughput gene-expression profiling technologies are now allowing multiplexed assessment of alterations responsible for the genesis and progression of bladder tumors. These investigations employ global or pathway-based approaches to define molecular signatures that can predict prognosis independent of traditional clinical performance metrics. Prognostic panels generated using these strategies can also elucidate the biology of tumor progression and identify potential therapeutic targets.
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Affiliation(s)
- Georg Bartsch
- Institute of Urology, University of Southern California Keck School of Medicine, Los Angeles, CA 90033, USA
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9
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Du C, Wang Y. The immunoregulatory mechanisms of carcinoma for its survival and development. JOURNAL OF EXPERIMENTAL & CLINICAL CANCER RESEARCH : CR 2011; 30:12. [PMID: 21255410 PMCID: PMC3031251 DOI: 10.1186/1756-9966-30-12] [Citation(s) in RCA: 44] [Impact Index Per Article: 3.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 11/15/2010] [Accepted: 01/21/2011] [Indexed: 12/24/2022]
Abstract
The immune system in patients detects and eliminates tumor cells, but tumors still progress persistently. The mechanisms by which tumor cells survive under the pressure of immune surveillance are not fully understood. This review is to present the evidence from clinical studies, showing a significant correlation of clinicopathological features of carcinoma with: (1) the loss of classical human leukocyte antigen class I, (2) the up-regulation of non-classical human leukocyte antigen class I, pro-apoptotic Fas ligand and receptor-binding cancer antigen expressed on SiSo cells I, and (3) the formation of immunosuppressive microenvironment by up-regulation of transforming growth factor-beta, Galectin-1, inhibitory ligand B7s, indoleamine 2,3-dioxygenase and arginase, as well as by recruitment of tumor-induced myeloid-derived suppressor cells and regulatory T cells. All of these factors may together protect carcinoma cells from the immune-cytotoxicity.
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Affiliation(s)
- Caigan Du
- Department of Urologic Sciences, University of British Columbia, Vancouver, Canada.
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Mitra AP, Bartsch CC, Cote RJ. Strategies for molecular expression profiling in bladder cancer. Cancer Metastasis Rev 2010; 28:317-26. [PMID: 19997771 DOI: 10.1007/s10555-009-9196-5] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/27/2023]
Abstract
Carcinoma of the urinary bladder involves alterations in multiple cellular pathways that dictate the pathology of the disease and clinical outcome of the patient. This includes alterations in regulation of the cell cycle, apoptotic mechanisms, signal transduction and tumor angiogenesis. Interrogation of alterations in multiple molecules associated with these pathways is leading to the development of biomarker panels that are capable of predicting an individual patient's outcome or response to specific treatments. With respect to gene expression profiling, two broad approaches may be identified: a global approach and a pathway-specific approach. The global approach involves a high-throughput effort to profile the entire genome, while the pathway-specific approach quantifies select genes across several pathways. While the former has a high potential for discovery of novel signatures, the latter is important in generating reproducible and concise panels that have the potential for rapid clinical implementation. A combination of both these approaches is needed for the identification and validation of robust marker panels of potential clinical importance in bladder cancer.
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Affiliation(s)
- Anirban P Mitra
- Department of Pathology, University of Southern California Keck School of Medicine, 2011 Zonal Avenue, HMR 308, Los Angeles, CA 90033, USA.
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Mitra AP, Cote RJ. Molecular pathogenesis and diagnostics of bladder cancer. ANNUAL REVIEW OF PATHOLOGY-MECHANISMS OF DISEASE 2009; 4:251-85. [PMID: 18840072 DOI: 10.1146/annurev.pathol.4.110807.092230] [Citation(s) in RCA: 150] [Impact Index Per Article: 9.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/09/2022]
Abstract
Despite elaborate characterization of the risk factors, bladder cancer is still a major epidemiological problem whose incidence continues to rise each year. Urothelial carcinoma is now recognized as a disease of alterations in several cellular processes. The more prevalent, less aggressive, recurrent, noninvasive tumors are characterized by constitutive activation of the Ras-MAPK pathway. The less common but more aggressive invasive tumors, which have a higher mortality rate, are characterized by alterations in the p53 and retinoblastoma pathways. Several diagnostic tests have attempted to identify these molecular alterations in tumor cells exfoliated in the urine, whereas prognostic tests have tried to identify aberrations so as to predict tumor behavior and identify therapeutic targets. The future of bladder cancer patient management will rely on the use of molecular tests to reliably diagnose the presence of disease, predict individual tumor behavior, and suggest potential targeted therapeutics.
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Affiliation(s)
- Anirban P Mitra
- Department of Pathology, Keck School of Medicine and Norris Comprehensive Cancer Center, University of Southern California, Los Angeles, CA 90033, USA.
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12
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Ellinger J, Bastian PJ, Ellinger N, Kahl P, Perabo FG, Büttner R, Müller SC, Ruecker AV. Apoptotic DNA fragments in serum of patients with muscle invasive bladder cancer: a prognostic entity. Cancer Lett 2008; 264:274-80. [PMID: 18329789 DOI: 10.1016/j.canlet.2008.01.038] [Citation(s) in RCA: 47] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/28/2007] [Revised: 01/21/2008] [Accepted: 01/23/2008] [Indexed: 11/18/2022]
Abstract
Patients with malignancies often possess increased concentrations of cell-free serum DNA. In this study, we investigated serum DNA levels in each 45 patients with bladder cancer (BCA) undergoing radical cystectomy and with benign prostate hyperplasia. A quantitative real-time PCR was used to amplify a 124 bp (PTGS2; mostly apoptotic origin) and a 271 bp (Reprimo; mostly necrotic origin) DNA fragment. Changes in the origin of DNA fragments were specified as the Apoptosis Index (AI, ratio of 124 bp/271 bp fragments). Small and large fragments were increased (p<0.001 and p=0.041) in BCA patients. The AI increase suggests that DNA fragmentation was mostly (p<0.001) caused by apoptosis. High levels of small DNA fragments distinguished between BCA and BPH with high sensitivity (96%) and moderate specificity (62%). DNA levels and the AI were not correlated with clinicopathological parameters. However, an increased AI was correlated with BCA-specific mortality in a multivariate analysis (p=0.011) indicating that the AI is an independent prognostic factor. Thus, cell-free DNA seems to be a useful prognostic marker in patients with BCA.
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Affiliation(s)
- Jörg Ellinger
- Klinik und Poliklinik für Urologie, Universitätsklinikum Bonn, Sigmund-Freud-Strasse 25, 53105 Bonn, Germany.
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Mao H, Liu Q, Zhang J, Gu H, Wang L, Zhou X, Yin H, Zhang L, Xie F, Jiang G. Effects of specific antisense oligonucleotide inhibition of Fas expression on T cell apoptosis induced by Fas ligand. Int Immunopharmacol 2007; 7:1714-22. [PMID: 17996681 DOI: 10.1016/j.intimp.2007.09.010] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/08/2007] [Revised: 08/21/2007] [Accepted: 09/05/2007] [Indexed: 01/15/2023]
Abstract
To investigate the effect of specific antisense oligodeoxynucleotide (ASODN) inhibition of Fas expression on T cell apoptosis induced by hepatocarcinoma cells. Fas receptor (Fas) and Fas ligand (FasL) expressed by the hepatocarcinoma cell line HepG2.2.15 and Jurkat T cells were detected by flow cytometry (FCM) and the ability of FasL-inducing T cell apoptosis was tested by co-culture assay in vitro with HepG2.2.15 cells and Jurkat T cells. The Jurkat cells were transfected with Fas-ASODN using lipofectin, and the effects of Fas-ASODN on Fas mRNA level, Fas expression on T cells surface, and apoptosis were investigated by RT-PCR, FCM and co-culture assay, respectively. It was found that HepG2.2.15 cells expressing functional FasL could induce the apoptosis of Jurkat cells as demonstrated by co-culture assays. After the Jurkat cells were transfected with Fas ASODN, the level of Fas mRNA, the expression rate of Fas and the apoptotic rate induced by hepatocarcinoma cells were all decreased. As a conclusion, it is evident that hepatocarcinoma cells expressing FasL can induce apoptosis in Fas-expressing T cells, indicating that transfection of Fas ASODN can partially convert the immune inhibitory condition induced by hepatocarcinoma cells.
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Affiliation(s)
- Haiting Mao
- Institute of Immunology, Shandong University School of Medicine, Jinan, 250012, People's Republic of China.
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Li C, Wu W, Liu J, Qian L, Li A, Yang K, Wei Q, Zhou J, Zhang Z. Functional polymorphisms in the promoter regions of the FAS and FAS ligand genes and risk of bladder cancer in south China: a case–control analysis. Pharmacogenet Genomics 2006; 16:245-51. [PMID: 16538171 DOI: 10.1097/01.fpc.0000194425.58511.a7] [Citation(s) in RCA: 43] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
Abstract
BACKGROUND FAS and FASLG together initiate apoptosis, which prevents tumor development. FAS and FASLG polymorphisms in the promoter regions can alter the transcriptional activities and thus alter risk of cancer. We hypothesized that the FAS -1377G>A, -670A>G, and FASLG -844T>C polymorphisms are associated with risk of bladder cancer. METHODS In a hospital-based case-control study of 216 case patients with newly diagnosed bladder transitional cell carcinoma and 252 cancer-free controls frequency-matched by age and sex, we genotyped polymorphisms using PCR-restriction fragment length polymorphism. RESULTS We found a statistically significantly increased risk of bladder cancer associated with the FASLG -844CC genotype [adjusted OR=1.51; 95% CI 1.03-2.23] compared with -844 (CT+TT). Consistently, the FAS haplotype genotypes with 2-4 variant (risk) alleles (-1377A and -670A) were associated with an increased risk of bladder cancer compared to 0-1 variants (OR=2.14; 95% CI 1.10-4.16). Furthermore, when we evaluated these three polymorphisms together, we found that the combined genotypes with 4-6 variant (risk) alleles (-1377A, -670A, and -844C) were associated with an increased risk of bladder cancer (OR=1.58; 95% CI 1.07-2.34) compared to 1-3 variants, and this increased risk was more pronounced among subgroups of aged >50 years (OR=1.70; 95% CI 1.11-2.61) and smokers (OR=1.88; 95% CI 1.06-3.32). CONCLUSIONS FAS and FASLG polymorphisms appear to jointly contribute to risk of bladder cancer in this southern Chinese population. Larger studies are needed to verify these findings.
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Affiliation(s)
- Chunping Li
- Department of Molecular Genetic Toxicology, Nanjing Medical University, 140 Hanzhong Road, Nanjing 210029, China
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15
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O'Kane HF, Watson CJ, Johnston SR, Petak I, Watson RWG, Williamson KE. Targeting death receptors in bladder, prostate and renal cancer. J Urol 2006; 175:432-8. [PMID: 16406966 DOI: 10.1016/s0022-5347(05)00160-6] [Citation(s) in RCA: 31] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/13/2005] [Indexed: 12/24/2022]
Abstract
PURPOSE We describe key components of normal and aberrant death receptor pathways, the association of these abnormalities with tumorigenesis in bladder, prostate and renal cancer, and their potential application in novel therapeutic strategies targeted toward patients with cancer. MATERIALS AND METHODS A MEDLINE literature search of the key words death receptors, TRAIL (tumor necrosis factor related apoptosis inducing ligand), FAS, bladder, prostate, renal and cancer was done to obtain information for review. A brief overview of the TRAIL and FAS death receptor pathways, and their relationship to apoptosis is described. Mechanisms that lead to nonfunction of these pathways and how they may contribute to tumorigenesis are linked. Current efforts to target death receptor pathways as a therapeutic strategy are highlighted. RESULTS Activation of tumor cell expressing death receptors by cytotoxic immune cells is the main mechanism by which the immune system eliminates malignant cells. Death receptor triggering induces a caspase cascade, leading to tumor cell apoptosis. Receptor gene mutation or hypermethylation, decoy receptor or splice variant over expression, and downstream inhibitor interference are examples of the ways that normal pathway functioning is lost in cancers of the bladder and prostate. Targeting death receptors directly through synthetic ligand administration and blocking downstream inhibitor molecules with siRNA or antisense oligonucleotides represent novel therapeutic strategies under development. CONCLUSIONS Research into the death receptor pathways has demonstrated the key role that pathway aberrations have in the initiation and progression of malignancies of the bladder, prostate and kidney. This new understanding has resulted in exciting approaches to restore the functionality of these pathways as a novel therapeutic strategy.
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Affiliation(s)
- Hugh F O'Kane
- Uro-oncology Group, Queen's University Belfast, Belfast, United Kingdom.
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16
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Zhu Q, Liu JY, Xu HW, Yang CM, Zhang AZ, Cui Y, Wang HB. Mechanism of counterattack of colorectal cancer cell by Fas/Fas ligand system. World J Gastroenterol 2005; 11:6125-9. [PMID: 16273638 PMCID: PMC4436628 DOI: 10.3748/wjg.v11.i39.6125] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: To determine the role of Fas/Fas ligand (FasL) in the immune escape of colon cancer cells.
METHODS: Immunohistochemistry was used to observe the expression of Fas and FasL in the tissues of colon cancer patients.In situ hybridization was used to detect the localization of FasL mRNA expression in cancer tissues. Terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL) assay and CD45 staining were performed to detect the apoptosis of tumor-infiltrating lymphocytes (TILs). Co-culture assays of colon cancer cells (SW480) and Jurkat cells (Fas-sensitive cells) were performed to observe the counterattack of colon cancer cells to lymphocytes.
RESULTS: Of 53 cases of colon carcinomas, 23 cases (43.4%) expressed Fas which was significantly lower as compared to the normal colonic mucosa (73.3%, P<0.01), and 45 cases (84.9%) of colon carcinomas expressed FasL, whereas only two cases (3.75%) in normal mucosa expressed FasL. FasL expression in the colon cancer cells was found to be associated with increased cell death of TILs. The apoptotic rate of TIL in the FasL-positive staining regions of tumor cells was significantly higher than that in the FasL-negative staining region (54.84 ± 2.79% vs 25.73 ± 1.98%, P<0.01). The co-culture of SW480 cells and Jurkat cells confirmed the function of FasL on the SW480 cells. The apoptotic rates of Jurkat cells were found to be related with the amount of SW480 cells.
CONCLUSION: Colon cancer cells can escape the immune surveillance and killing via decreasing Fas expression, and can counterattack the immune system via increasing FasL expression. Fas/FasL can serve as potential targets for effective antitumor therapy.
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Affiliation(s)
- Qiang Zhu
- Department of Gastroenterology, Shandong Provincial Hospital, Jing 5 Wei 7 Road, 324#, Jinan 250021, Shandong Province, China.
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17
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Abstract
The role of the immune system in controlling the growth of tumour cells is highly complex and has been extensively debated. It is well documented that the immune system controls virally induced cancers, and there is evidence for a role of specific immunity in other types of tumours. The greater understanding of the regulation and optimization of adoptive, specific immune responses, and the better characterization of tumour-associated antigens indicate the way for active specific vaccination and cell therapy in urological tumours. Currently, bacille Calmette Guerin immunotherapy is established for localized bladder cancer and many experimental immunotherapies are under evaluation. Here we review some timely aspects of tumour immunology, and describe the current status and development of immunotherapy in prostate and bladder cancer.
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Affiliation(s)
- Thomas H Totterman
- Clinical Immunology Division, Department of Oncology, Radiology and Clinical Immunology, Rudbeck Laboratory, Uppsala University Hospital, Uppsala, Sweden
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18
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Li M, Liu X, Zhou S, Li P, Li G. Effects of alpha fetoprotein on escape of Bel 7402 cells from attack of lymphocytes. BMC Cancer 2005; 5:96. [PMID: 16080799 PMCID: PMC1198224 DOI: 10.1186/1471-2407-5-96] [Citation(s) in RCA: 36] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/08/2005] [Accepted: 08/05/2005] [Indexed: 12/15/2022] Open
Abstract
BACKGROUND Involvement of AFP against apoptosis of tumor cell has been implicated in its evasion of immune surveillance. However, the molecular events of immune escape mechanisms are still unknown. The major observations reported here relate to a possible mechanism by which heptoloma Bel 7402 cells escape immune surveillance in vitro. METHODS Western blotting and a well-characterized cofocal scanning image were performed to analyze the expression of Fas/FasL and caspase-3 in co-cultured Bel 7402 and Jurkat cells. RESULTS After co-culture with Jurkat cells, up-regulated Fas and reduced FasL expression could be observed. Treatment with AFP could remarkably inhibit the elevated Fas and, whereas, induce the FasL expression in co-cultured Bel 7402 cells. Cells co-culture could induce the expression of caspase-3 in both cells line. The elevated caspase-3 in Bel 7402 cells was abolished following the treatment of AFP. The expression of caspase-3 was elevated in co-cultured Jurkat cells treated with AFP. No detectable change on the expression of survivin was examined in both cells line. Monoclonal antibody against AFP treatment alone did not obviously influence the growth of cells, as well as the expression of Fas/FasL and caspase-3. However, the effect of AFP could be blocked by antibody. CONCLUSIONS our results provide evidence that AFP could promote the escape of liver cancer cells from immune surveillance through blocking the caspase signal pathway of tumor cells and triggering the Fas/FasL interaction between tumor cells and lymphocytes.
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Affiliation(s)
- Mengsen Li
- Department of Biochemistry and Molecular Biology, Peking University Health Science Center, Beijing 100083, China
| | - Xinhua Liu
- Department of Biochemistry and Molecular Biology, Peking University Health Science Center, Beijing 100083, China
| | - Sheng Zhou
- Department of Biochemistry, Hainan Medical College, Haikou 570102, China
| | - Pingfeng Li
- Department of Biochemistry and Molecular Biology, Peking University Health Science Center, Beijing 100083, China
| | - Gang Li
- Department of Biochemistry and Molecular Biology, Peking University Health Science Center, Beijing 100083, China
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19
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Li M, Liu X, Zhou S, Li P, Li G. Effects of alpha fetoprotein on escape of Bel 7402 cells from attack of lymphocytes. BMC Cancer 2005. [PMID: 16080799 DOI: 10.1186/1471-2407-5-96|issn] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/10/2022] Open
Abstract
BACKGROUND Involvement of AFP against apoptosis of tumor cell has been implicated in its evasion of immune surveillance. However, the molecular events of immune escape mechanisms are still unknown. The major observations reported here relate to a possible mechanism by which heptoloma Bel 7402 cells escape immune surveillance in vitro. METHODS Western blotting and a well-characterized cofocal scanning image were performed to analyze the expression of Fas/FasL and caspase-3 in co-cultured Bel 7402 and Jurkat cells. RESULTS After co-culture with Jurkat cells, up-regulated Fas and reduced FasL expression could be observed. Treatment with AFP could remarkably inhibit the elevated Fas and, whereas, induce the FasL expression in co-cultured Bel 7402 cells. Cells co-culture could induce the expression of caspase-3 in both cells line. The elevated caspase-3 in Bel 7402 cells was abolished following the treatment of AFP. The expression of caspase-3 was elevated in co-cultured Jurkat cells treated with AFP. No detectable change on the expression of survivin was examined in both cells line. Monoclonal antibody against AFP treatment alone did not obviously influence the growth of cells, as well as the expression of Fas/FasL and caspase-3. However, the effect of AFP could be blocked by antibody. CONCLUSIONS our results provide evidence that AFP could promote the escape of liver cancer cells from immune surveillance through blocking the caspase signal pathway of tumor cells and triggering the Fas/FasL interaction between tumor cells and lymphocytes.
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Affiliation(s)
- Mengsen Li
- Department of Biochemistry and Molecular Biology, Peking University Health Science Center, Beijing 100083, China.
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20
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Sudarshan S, Holman DH, Hyer ML, Voelkel-Johnson C, Dong JY, Norris JS. In vitro efficacy of Fas ligand gene therapy for the treatment of bladder cancer. Cancer Gene Ther 2005; 12:12-8. [PMID: 15514684 DOI: 10.1038/sj.cgt.7700746] [Citation(s) in RCA: 37] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/05/2023]
Abstract
Previous investigations have revealed that bladder cancer cells are generally resistant to Fas-mediated apoptosis by conventional Fas agonists. However, the ability of these cell lines to undergo Fas-mediated apoptosis may have been underappreciated. As a result, we investigated the in vitro efficacy of Fas ligand gene therapy for bladder cancer. Three human bladder cancer lines (T24, J82, and 5637) were treated with the conventional Fas agonist CH-11, a monoclonal antibody to the Fas receptor. Cells were also treated with a replication-deficient adenovirus containing a modified murine Fas ligand gene fused to green fluorescent protein (GFP), AdGFPFasL. A virus containing the GFP gene alone was used to control for viral toxicity (AdGFP). Cell death was quantified using a tetrazolium-based (MTS) assay. Cells were also evaluated by Western blotting to evaluate poly (ADP-ribose) polymerase, caspase 8, and caspase 9 cleavage and by flow cytometry to determine the presence of coxsackie/adenovirus receptor (CAR). These studies confirmed bladder cancer resistance to cell death by the anti-Fas monoclonal antibody CH-11. This resistance was overcome with AdGFPFasL at a multiplicity of infection (MOI) of 1000 achieving over 80% cell death in all cell lines. Furthermore, greater than 80% cell death was evident in 5637 cells treated with low-dose AdGFPFasL (MOI=10). 5637 cells expressed significantly higher levels of surface CAR than J82 or T24 cells (P<.05). AdGFPFasL is cytotoxic to bladder cancer cells that would otherwise be considered Fas resistant, supporting its in vivo potential. Enhanced sensitivity to AdGFPFasL may be in part due to increased cell surface CAR levels.
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Affiliation(s)
- Sunil Sudarshan
- Department of Microbiology and Immunology, Medical University of South Carolina, Charleston, South Carolina, USA
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21
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Korkolopoulou P, Goudopoulou A, Voutsinas G, Thomas-Tsagli E, Kapralos P, Patsouris E, Saetta AA. c-FLIP expression in bladder urothelial carcinomas: its role in resistance to Fas-mediated apoptosis and clinicopathologic correlations. Urology 2004; 63:1198-204. [PMID: 15183989 DOI: 10.1016/j.urology.2004.01.007] [Citation(s) in RCA: 55] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/10/2003] [Accepted: 01/07/2004] [Indexed: 11/30/2022]
Abstract
OBJECTIVES To investigate the incidence of Fas (exon 9) mutations and the expression of Fas, Fas-Fas ligand (FasL) system, and cellular FLICE-like inhibitory protein (c-FLIP) in relation to standard clinicopathologic parameters and patient outcome in bladder carcinoma. Disruption of apoptotic cell death has been implicated in tumor aggressiveness in bladder urothelial carcinomas. The FasL system is involved in the execution of apoptosis induced by the immune system. c-FLIP protein constitutes an important endogenous inhibitor of Fas and other death receptor-mediated apoptosis. METHODS The expression of Fas, FasL, and c-FLIP was quantified immunohistochemically in paraffin-embedded tissues from 53 patients for whom clinical information was available. DNA extracted from the same samples was screened for mutations in Fas exon 9 by single-strand conformation polymorphism and sequencing. The effect of Fas, FasL, and c-FLIP on clinical outcome was assessed by univariate and multivariate analyses. RESULTS Positive immunostaining was detected for Fas, FasL, and c-FLIP in 72%, 66%, and 81% of cases, respectively. Concurrent expression of Fas and FasL was seen in 27 samples (51%), of which 22 (81.5%) also displayed c-FLIP positivity. FasL and c-FLIP expression increased with advancing stage but was absent from normal urothelium. None of the 53 urothelial carcinoma samples analyzed showed evidence of mutations by polymerase chain reaction single-strand conformation polymorphism and direct sequencing. Survival analysis demonstrated that although both FasL and c-FLIP expression adversely affected survival, only c-FLIP remained statistically significant on multivariate analysis. CONCLUSIONS The frequent expression and coexpression of Fas, FasL, and c-FLIP in urothelial carcinomas implicates c-FLIP as an inhibitor of the Fas-FasL-induced death pathway in these tumors. Moreover, c-FLIP conveys independent prognostic information in the presence of classical prognosticators.
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Affiliation(s)
- Penelope Korkolopoulou
- Department of Pathology, National and Kapodistrian University of Athens Medical School, Athens, Greece
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22
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Filion MC, Filion B, Roy J, Ménard S, Reader S, Phillips NC. Development of immunomodulatory six base-length non-CpG motif oligonucleotides for cancer vaccination. Vaccine 2004; 22:2480-8. [PMID: 15193412 DOI: 10.1016/j.vaccine.2003.11.072] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/26/2003] [Accepted: 11/18/2003] [Indexed: 01/05/2023]
Abstract
We have previously described a novel family of immunomodulatory synthetic oligonucleotides characterized by a phosphodiester backbone, a length of six bases and a 5'G3xG23' sequence, where x is A, C, G or T. In the present study, we have evaluated whether these 5'G3xG23' oligonucleotides possess additional activities essential for adequate cancer vaccination. Immunization for the treatment of cancer requires an adjuvant, a source of tumor-associated antigen(s), for example apoptotic cancer cells, and a way to overcome the escape of tumor cells from the immune system, for example the up-regulation of Fas ligand (FasL) on the surface of cancer cells. The results show that phosphodiester 5'G3AG23' and 5'G3TG23' oligonucleotides have a direct activity on a number of different cancer cells by inducing apoptosis (release of cytochrome C, activation of caspase-3, cleavage of poly [ADP-ribose] polymerase, degradation of nuclear mitotic apparatus protein and translocation of phophatidylserine at the cell surface). In addition, the 5'G3AG23', 5'G3CG23', and 5'G3TG23' oligonucleotides were found to down-regulate the levels of FasL on the surface of cancer cells. These immunomodulatory phosphodiester six base-length oligonucleotides, which are capable of inducing apoptosis in cancer cells as well as downregulating the expression of FasL at their cell surface, may have application as cancer cell vaccines.
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Affiliation(s)
- Mario C Filion
- Bioniche Therapeutics Division, Bioniche Life Sciences, 6100 Royalmount Avenue, Montréal, Que., Canada, H4P 2R2.
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23
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Maas S, Warskulat U, Steinhoff C, Mueller W, Grimm MO, Schulz WA, Seifert HH. Decreased Fas expression in advanced-stage bladder cancer is not related to p53 status. Urology 2004; 63:392-7. [PMID: 14972509 DOI: 10.1016/j.urology.2003.08.023] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/14/2003] [Accepted: 08/22/2003] [Indexed: 10/26/2022]
Abstract
OBJECTIVES The Fas-Fas ligand system is an important regulator of apoptosis and is involved in tumor development. Invasive cancers downregulate Fas expression to evade antitumor immune responses. Fas is a transcriptional target of p53, which is often mutated in bladder cancers. Therefore, Fas expression and its relation to p53 mutation was investigated. METHODS Expression of Fas protein and p53 status was studied by immunohistochemistry in 83 bladder cancer specimens. In addition, mRNA levels for soluble (decoy) and membrane-bound forms of Fas were compared between 10 bladder cancer cell lines and primary uroepithelial cells by quantitative TaqMan polymerase chain reaction. Mutational analysis of the death domain of the Fas gene was performed in all cell lines. RESULTS Organ-confined tumors maintained specific Fas staining at the cell membrane and often also in the cytoplasm. In higher stage carcinomas, Fas expression became restricted to a smaller fraction of cells or was lacking entirely. The correlation of Fas staining with tumor stage was highly significant but no correlation to tumor grade or survival was found. Furthermore, no statistically significant relationship was observed with either the presence or lack of mutated p53 accumulation. Membrane-bound Fas mRNA was decreased in most, and soluble Fas was increased in all transitional cell carcinoma lines compared with primary uroepithelial cells. No mutations in the death domain were detected. CONCLUSIONS Fas downregulation occurring in advanced bladder cancer is unrelated to p53 mutations. The results of immunohistochemistry and mRNA studies of soluble and membrane-bound Fas in transitional cell carcinoma lines support the hypothesis of immune evasion in advanced bladder cancer.
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MESH Headings
- Carcinoma, Papillary/genetics
- Carcinoma, Papillary/metabolism
- Carcinoma, Papillary/pathology
- Carcinoma, Transitional Cell/genetics
- Carcinoma, Transitional Cell/metabolism
- Carcinoma, Transitional Cell/mortality
- Carcinoma, Transitional Cell/pathology
- Cell Line, Tumor/metabolism
- Cell Membrane/metabolism
- Down-Regulation
- Gene Expression Regulation, Neoplastic
- Genes, p53
- Humans
- Life Tables
- Neoplasm Proteins/biosynthesis
- Neoplasm Proteins/genetics
- Neoplasm Staging
- Polymorphism, Single-Stranded Conformational
- RNA, Messenger/biosynthesis
- RNA, Neoplasm/biosynthesis
- Solubility
- Survival Analysis
- Tumor Escape/genetics
- Urinary Bladder Neoplasms/genetics
- Urinary Bladder Neoplasms/metabolism
- Urinary Bladder Neoplasms/mortality
- Urinary Bladder Neoplasms/pathology
- fas Receptor/biosynthesis
- fas Receptor/genetics
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Affiliation(s)
- Simone Maas
- Department of Urology, Heinrich-Heine-University, Duesseldorf, Germany
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24
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Loskog A, Dzojic H, Vikman S, Ninalga C, Essand M, Korsgren O, Totterman TH. Adenovirus CD40 Ligand Gene Therapy Counteracts Immune Escape Mechanisms in the Tumor Microenvironment. THE JOURNAL OF IMMUNOLOGY 2004; 172:7200-5. [PMID: 15153545 DOI: 10.4049/jimmunol.172.11.7200] [Citation(s) in RCA: 62] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/19/2022]
Abstract
Tumors exhibit immune escape properties that promote their survival. These properties include modulation of Ag presentation, secretion of immunosuppressive factors, resistance to apoptosis, and induction of immune deviation, e.g., shifting from Th1- to Th2-type responses. These escape mechanisms have proven to hamper several immunotherapeutic strategies, and efforts need to be taken to revert this situation. We have studied the immunological effects of introducing CD40 ligand (CD40L), a potent dendritic cell activation molecule, into the tumor micromilieu by adenoviral gene transfer. For this purpose, a murine bladder cancer model (MB49) was used in C57BL/6 mice. The MB49 cells are known to induce IL-10 in the tumor environment. IL-10 potently inhibits the maturation of dendritic cells and thereby also the activation of CTLs. In this paper we show that CD40L immunogene therapy suppresses IL-10 and TGF-beta production (2-fold decrease) and induces a typical Th1-type response in the tumor area (200-fold increase in IL-12 production). The antitumor responses obtained were MB49 cell specific, and the cytotoxicity of the stimulated CD8(+) cells could be blocked by IL-10. Adenovirus CD40L therapy was capable of regressing small tumors (five of six animals were tumor free) and inhibiting the progression of larger tumors even in the presence of other escape mechanisms, such as apoptosis resistance. Furthermore, CD40L-transduced MB49 cells promoted the maturation of dendritic cells (2-fold increase in IL-12) independently of IL-10. Our results argue for using adenovirus CD40L gene transfer, alone or in combination with other modalities, for the treatment of Th2-dominated tumors.
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Affiliation(s)
- Angelica Loskog
- Clinical Immunology Division, Rudbeck Laboratory, Uppsala University, Uppsala, Sweden.
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25
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Abstract
Prostate cancer is the second leading cause of cancer death in the US, largely because of the limitations of our current therapeutic options, especially once the cancer has metastasized. Investigators have long sought new therapeutic modalities such as angiogenesis inhibitors, vaccines, and gene therapy, among others. It appears that a combination approach will be required to cure the majority of malignancies. Immunotherapy for prostate cancer appears feasible and a likely therapeutic modality in the armamentarium. Unfortunately, further research in basic immunology and the interaction of the immune system with other forms of therapy is needed. Many obstacles exist in immunotherapy, including vector design, tumouricidal specificity, and tumor evasion, which will have to be overcome in order to realize the maximum therapeutic benefit from this treatment modality.
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Affiliation(s)
- Joseph M Kaminski
- Department of Radiology, Medical College of Georgia, Augusta, GA 30912, USA
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26
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Chopin D, Barei-Moniri R, Maillé P, Le Frère-Belda MA, Muscatelli-Groux B, Merendino N, Lecerf L, Stoppacciaro A, Velotti F. Human urinary bladder transitional cell carcinomas acquire the functional Fas ligand during tumor progression. THE AMERICAN JOURNAL OF PATHOLOGY 2003; 162:1139-49. [PMID: 12651606 PMCID: PMC1851234 DOI: 10.1016/s0002-9440(10)63910-7] [Citation(s) in RCA: 25] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/20/2022]
Abstract
The interaction between FasL on tumor cells and Fas on lymphocytes may represent a tumor immune escape mechanism. We explored FasL expression and function in human urinary bladder transitional cell carcinomas (TCCs). FasL expression was observed in situ in 45% of TCCs (n = 45) and was absent in normal urothelium (n = 20). A correlation existed between FasL expression and high tumor grade (0% in G1, 14% in G2, and 75% in G3; P < 0.0001) and stage (13% in superficial Ta-T1 versus 81% in invasive T2-T4; P < 0.0001). FasL function was shown by the ability of two FasL-positive primary culture TCC cell lines (established from two FasL-positive invasive TCCs) to induce Fas-mediated killing not only of conventional Fas-sensitive targets (such as Jurkat cells or phytohemagglutinin-lymphoblasts), but also of autologous T lymphocytes generated in a mixed lymphocyte tumor-cell culture. In addition, an association between FasL expression by TCC cells and activated caspase-8, -9, and -3 expression by interferon-gamma-producing CD8-positive tumor-infiltrating lymphocytes was observed in situ. Our results show a functional expression of TCC-expressed FasL that correlates with tumor progression. These results suggest that TCC-expressed FasL may induce apoptosis of anti-tumor T lymphocytes in vivo, providing new insights on the mechanisms involved in bladder TCC progression.
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Affiliation(s)
- Dominique Chopin
- Service d'Urologie et Groupe d'Etude des Tumeurs Urologiques Equipe Mixte INSERM 03-37, Centre Hôspitalier Universitaire Henri Mondor, Créteil, France
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