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Lyu SY, Meshesha SM, Hong CE. Synergistic Effects of Mistletoe Lectin and Cisplatin on Triple-Negative Breast Cancer Cells: Insights from 2D and 3D In Vitro Models. Int J Mol Sci 2025; 26:366. [PMID: 39796221 PMCID: PMC11719730 DOI: 10.3390/ijms26010366] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/21/2024] [Revised: 12/09/2024] [Accepted: 12/30/2024] [Indexed: 01/13/2025] Open
Abstract
Triple-negative breast cancer (TNBC) remains a challenging subtype due to its aggressive nature and limited treatment options. This study investigated the potential synergistic effects of Korean mistletoe lectin (Viscum album L. var. coloratum agglutinin, VCA) and cisplatin on MDA-MB-231 TNBC cells using both 2D and 3D culture models. In 2D cultures, the combination of VCA and cisplatin synergistically inhibited cell proliferation, induced apoptosis, and arrested the cell cycle at the G2/M phase. Also, the combination treatment significantly reduced cell migration and invasion. Gene expression analysis showed significant changes in specific genes related to apoptosis (Bax, Bcl-2), metastasis (MMP-2, MMP-9), and EMT (E-cadherin, N-cadherin). Three-dimensional spheroid models corroborated these findings, demonstrating enhanced cytotoxicity and reduced invasion with the combination treatment. Significantly, the 3D models exhibited differential drug sensitivity compared to 2D cultures, emphasizing the importance of utilizing physiologically relevant models in preclinical studies. The combination treatment also reduced the expression of angiogenesis-related factors VEGF-A and HIF-1α. This comprehensive study provides substantial evidence for the potential of VCA and cisplatin combination therapy in TNBC treatment and underscores the significance of integrating 2D and 3D models in preclinical cancer research.
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Affiliation(s)
- Su-Yun Lyu
- College of Pharmacy, Sunchon National University, Suncheon 57922, Republic of Korea; (S.-Y.L.)
- Research Institute of Life and Pharmaceutical Sciences, Sunchon National University, Suncheon 57922, Republic of Korea
| | - Saporie Melaku Meshesha
- College of Pharmacy, Sunchon National University, Suncheon 57922, Republic of Korea; (S.-Y.L.)
| | - Chang-Eui Hong
- College of Pharmacy, Sunchon National University, Suncheon 57922, Republic of Korea; (S.-Y.L.)
- Research Institute of Life and Pharmaceutical Sciences, Sunchon National University, Suncheon 57922, Republic of Korea
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Robev B, Iliev I, Tsoneva I, Momchilova A, Nesheva A, Kostadinova A, Staneva G, Nikolova B. Antitumor Effect of Iscador on Breast Cancer Cell Lines with Different Metastatic Potential. Int J Mol Sci 2023; 24:ijms24065247. [PMID: 36982323 PMCID: PMC10049140 DOI: 10.3390/ijms24065247] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/23/2023] [Revised: 03/02/2023] [Accepted: 03/06/2023] [Indexed: 03/12/2023] Open
Abstract
Studies were performed for the first time on the effect of Iscador Qu and Iscador M on phototoxicity, cytotoxicity, antiproliferative activity, changes in ξ-potential of cells, membrane lipid order, actin cytoskeleton organization and migration on three breast cancer lines with different metastatic potential: MCF10A (control), MCF-7 (low metastatic) and MDA-MB231 (high metastatic) cells. The tested Iscador Qu and M did not show any phototoxicity. The antiproliferative effect of Iscador species appeared to be dose-dependent and was related to the metastatic potential of the tested cell lines. A higher selectivity index was obtained for Iscador Qu and M towards the low metastatic MCF-7 cell line compared to the high metastatic MDA-MB-231. Iscador Qu demonstrated higher selectivity for both cancer cell lines compared to Iscador M. The malignant cell lines exhibited a decrease in fibril number and thickness regardless of the type of Iscador used. The strongest effect on migration potential was observed for the low metastatic cancer cell line MCF-7 after Iscador treatment. Both Iscador species induced a slight increase in the percentage of cells in early apoptosis for the low and high metastatic cell lines, MCF-7 and MDA-MB-231, unlike control cells. Changes in the zeta potential and membrane lipid order were observed for the low metastatic MCF-7 cell line in contrast to the high metastatic MDA-MB-231 cells. The presented results reveal a higher potential of Iscador as an antitumor agent for the low metastatic cancer cell line MCF-7 compared to the high metastatic one. Iscador Qu appears to be more potent compared to Iscador M, but at this point, the exact mechanism of action is still unclear and needs further investigations.
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Affiliation(s)
- Bozhil Robev
- Department of Medical Oncology, University Hospital “Sv. Ivan Rilski”, 15 Acad. Ivan Geshov Blvd., 1431 Sofia, Bulgaria
| | - Ivan Iliev
- Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 25, 1113 Sofia, Bulgaria
| | - Iana Tsoneva
- Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 21, 1113 Sofia, Bulgaria
| | - Albena Momchilova
- Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 21, 1113 Sofia, Bulgaria
- Correspondence:
| | - Alexandrina Nesheva
- Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 21, 1113 Sofia, Bulgaria
| | - Aneliya Kostadinova
- Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 21, 1113 Sofia, Bulgaria
| | - Galya Staneva
- Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 21, 1113 Sofia, Bulgaria
| | - Biliana Nikolova
- Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 21, 1113 Sofia, Bulgaria
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EGFR and p38MAPK Contribute to the Apoptotic Effect of the Recombinant Lectin from Tepary Bean (Phaseolus acutifolius) in Colon Cancer Cells. Pharmaceuticals (Basel) 2023. [DOI: 10.3390/ph16020290] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/16/2023] Open
Abstract
Previous works showed that a Tepary bean lectin fraction (TBLF) induced apoptosis on colon cancer cells and inhibited early colonic tumorigenesis. One Tepary bean (TB) lectin was expressed in Pichia pastoris (rTBL-1), exhibiting similarities to one native lectin, where its molecular structure and in silico recognition of cancer-type N-glycoconjugates were confirmed. This work aimed to determine whether rTBL-1 retained its bioactive properties and if its apoptotic effect was related to EGFR pathways by studying its cytotoxic effect on colon cancer cells. Similar apoptotic effects of rTBL-1 with respect to TBLF were observed for cleaved PARP-1 and caspase 3, and cell cycle G0/G1 arrest and decreased S phase were observed for both treatments. Apoptosis induction on SW-480 cells was confirmed by testing HA2X, p53 phosphorylation, nuclear fragmentation, and apoptotic bodies. rTBL-1 increased EGFR phosphorylation but also its degradation by the lysosomal route. Phospho-p38 increased in a concentration- and time-dependent manner, matching apoptotic markers, and STAT1 showed activation after rTBL-1 treatment. The results show that part of the rTBL-1 mechanism of action is related to p38 MAPK signaling. Future work will focus further on the target molecules of this recombinant lectin against colon cancer.
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Gomes DC, Barros MR, Menezes TM, Neves JL, Paiva PMG, da Silva TG, Napoleão TH, Coriolano MC, Dos Santos Correia MT. A new lectin from the floral capitula of Egletes viscosa (EgviL): Biochemical and biophysical characterization and cytotoxicity to human cancer cells. Int J Biol Macromol 2020; 168:676-685. [PMID: 33220373 DOI: 10.1016/j.ijbiomac.2020.11.124] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/15/2020] [Revised: 11/12/2020] [Accepted: 11/17/2020] [Indexed: 10/23/2022]
Abstract
Egletes viscosa is a plant with therapeutic value due to its antibacterial, antinociceptive and gastroprotective properties. This study aimed to purify, characterize, and evaluate the cytotoxicity of a lectin (EgviL) from the floral capitula of E. viscosa. The lectin was isolated from saline extract through precipitation with ammonium sulfate followed by Sephadex G-75 chromatography. The molecular mass and isoelectric point (pI) of EgviL were determined as well as its temperature and pH stability. Physical-chemical parameters of interaction between EgviL and carbohydrates were investigated by fluorescence quenching and 1H nuclear magnetic resonance (NMR). Cytotoxicity was investigated against human peripheral blood mononuclear cells (PBMCs) and neoplastic cells. EgviL (28.8 kDa, pI 5.4) showed hemagglutinating activity stable towards heating until 60 °C and at the pH range 5.0-7.0. This lectin is able to interact through hydrophobic and electrostatic bonds with galactose and glucose, respectively. EgviL reduced the viability of PBMCs only at the highest concentration tested (100 μg/mL) while was toxic to Jurkat E6-1 cells with IC50 of 24.1 μg/mL,inducing apoptosis. In summary, EgviL is a galactose/glucose-binding protein with acidic character, stable to heating and with cytotoxic effect on leukemic cells.
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Affiliation(s)
- Dayane Correia Gomes
- Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil
| | - Marcela Rodrigues Barros
- Laboratório de Química Biológica, Departamento de Química Fundamental, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil
| | - Thaís Meira Menezes
- Laboratório de Química Biológica, Departamento de Química Fundamental, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil
| | - Jorge Luiz Neves
- Laboratório de Química Biológica, Departamento de Química Fundamental, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil
| | - Patrícia Maria Guedes Paiva
- Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil
| | - Teresinha Gonçalves da Silva
- Departamento de Antibióticos, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil
| | - Thiago Henrique Napoleão
- Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil.
| | - Marília Cavalcanti Coriolano
- Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil.
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Mishra R, Sharma S, Sharma RS, Singh S, Sardesai MM, Sharma S, Mishra V. Viscum articulatum Burm. f. aqueous extract exerts antiproliferative effect and induces cell cycle arrest and apoptosis in leukemia cells. JOURNAL OF ETHNOPHARMACOLOGY 2018; 219:91-102. [PMID: 29555410 DOI: 10.1016/j.jep.2018.03.005] [Citation(s) in RCA: 17] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 07/20/2017] [Revised: 03/06/2018] [Accepted: 03/06/2018] [Indexed: 06/08/2023]
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE Viscum articulatum Burm. f. (leafless mistletoe) has been used in traditional system of medicines in India, China, Taiwan, Cambodia, Laos, and Vietnam, to treat blood-related diseases and various inflammatory and degenerative diseases including cancer. Anticancer activities of some phytomolecules purified from Viscum articulatum Burm. f. have been tested. However scientific evidence for the anticancerous potential of aqueous extract of V. articularum (VAQE) used in traditional medicine is lacking. AIM OF THE STUDY To study the antiproliferative and apoptotic effect of VAQE on Jurkat E6.1 and THP1 leukemia cells. MATERIALS AND METHODS The aqueous extract of the whole plant of Viscum articulatum Burm. f. was prepared in phosphate buffer saline. In VAQE, total soluble protein was estimated using Bradford's dye-binding assay; flavonoid content was determined using aluminum chloride colorimetric assay; and phenolic content was estimated following Folin-Ciocalteu colorimetric assay. XTT cell viability assay was used to test VAQE induced cytotoxicity in Jurkat E6.1 and THP1 leukemia cells and peripheral blood mononuclear cells (PBMC). The effect of VAQE on cell cycle progression was analyzed by PI staining using flow cytometry. Annexin-V-FITC/PI differential staining method was used for detecting the onset of apoptosis in leukemia cells. Rhodamine 123 dye was used to detect the change in mitochondrial membrane potential (MMP) using flow cytometry. DCF-DA fluorescence dye was used to estimate the level of reactive oxygen species (ROS). The ROS inhibitors were used to evaluate the role of ROS in mediating DNA degradation in VAQE-treated leukemia cells. The molecular mechanisms underlying VAQE induced apoptosis induction was studied by analyzing the expression of anti-apoptotic (Bcl-2) and pro-apoptotic (Bax) proteins, caspase-8 and caspase-3 enzymes using western blot. Diphenylamine (DPA) assay was used to determine the DNA fragmentation and conclusion of apoptosis. RESULTS VAQE triggered cytotoxic effect on Jurkat E6.1 (IC50-2.4 µg/ml; 24 h) and THP1 (IC50-1.0 µg/ml; 24 h) cells in a dose- and time-dependent manner. The apoptosis induction and G2/M arrest of the cell cycle are the cause of VAQE-induced cytotoxicity in leukemia cells. The apoptosis in VAQE-treated Jurkat E6.1 and THP1 cells was mediated via a reduction in MMP, elevation of intracellular ROS, decreased expression of the anti-apoptotic (Bcl-2) and increased expression of the pro-apoptotic (Bax) protein, activation of caspase-8 and caspase-3 and DNA fragmentation. CONCLUSION VAQE has a high efficacy to exert a cytotoxic effect in Jurkat E6.1 and THP1 cells and to induce apoptosis and G2/M cell cycle arrest. VAQE induces extrinsic pathway of apoptosis in both the leukemia cell lines via disruption of MMP, intracellular ROS imbalance, increased ratio of Bax/Bcl-2, activation of caspase-8, caspase-3 and ROS-mediated DNA fragmentation. The knowledge gained from the outcomes of the study may encourage the identification of novel chemotherapeutic agent from Viscum articulatum Burm. f. to treat leukemia.
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Affiliation(s)
- Ruchi Mishra
- Bioresources and Environmental Biotechnology Laboratory, Department of Environmental Studies, University of Delhi, Delhi 110007, India
| | - Saurabh Sharma
- Bioresources and Environmental Biotechnology Laboratory, Department of Environmental Studies, University of Delhi, Delhi 110007, India
| | - Radhey Shyam Sharma
- Bioresources and Environmental Biotechnology Laboratory, Department of Environmental Studies, University of Delhi, Delhi 110007, India
| | - Savita Singh
- Bioresources and Environmental Biotechnology Laboratory, Department of Environmental Studies, University of Delhi, Delhi 110007, India
| | | | - Sadhna Sharma
- Department of Zoology, Miranda House, University of Delhi, Delhi 110007, India
| | - Vandana Mishra
- Bioresources and Environmental Biotechnology Laboratory, Department of Environmental Studies, University of Delhi, Delhi 110007, India.
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Moringa oleifera seed lectin inhibits Ehrlich ascites carcinoma cell growth by inducing apoptosis through the regulation of Bak and NF-κB gene expression. Int J Biol Macromol 2018; 107:1936-1944. [DOI: 10.1016/j.ijbiomac.2017.10.070] [Citation(s) in RCA: 29] [Impact Index Per Article: 4.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/10/2017] [Revised: 10/10/2017] [Accepted: 10/11/2017] [Indexed: 01/22/2023]
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Articulatin-D induces apoptosis via activation of caspase-8 in acute T-cell leukemia cell line. Mol Cell Biochem 2016; 426:87-99. [DOI: 10.1007/s11010-016-2883-y] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/20/2016] [Accepted: 11/07/2016] [Indexed: 12/14/2022]
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Anti-cancer effects of enteric-coated polymers containing mistletoe lectin in murine melanoma cells in vitro and in vivo. Mol Cell Biochem 2015; 408:73-87. [PMID: 26152904 DOI: 10.1007/s11010-015-2484-1] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/30/2015] [Accepted: 06/18/2015] [Indexed: 02/03/2023]
Abstract
In this study, we evaluated the effects of Korean mistletoe (Viscum album L. var. coloratum) coated with a biodegradable polymer (Eudragit(®)) wall on the growth of mouse melanoma in vivo. Oral administration of 4% (430 mg/kg/day) enteric-coated mistletoe resulted in a significant reduction in tumor volume on day 14 compared to the negative control group in B16F10 melanoma-inoculated BDF1 mice. When we measured the survival rate, enteric-coated mistletoe-received mice had a higher survival rate after day 12. Also, we investigated the mechanism involving the cancer cell growth inhibition when melanoma cells were treated with Korean mistletoe lectin (Viscum album L. var. coloratum agglutinin, VCA) and its extract in vitro. As a result, a significant G0/G1 arrest was observed in both B16BL6 and B16F10 melanoma cells with VCA or mistletoe extract. In addition, VCA or mistletoe extract induced an increase in both early and late apoptosis in cells. When we studied the molecular mechanism, our results showed that VCA and mistletoe extract can increase activated multiple caspases (caspase-1, 3, 4, 5, 6, 7, 8, and 9), dose-dependently. We also found out that VCA and mistletoe treatment causes a significant decrease in the expression of procaspase-3 and 8.
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Lyu SY, Choi JH, Lee HJ, Park WB, Kim GJ. Korean mistletoe lectin promotes proliferation and invasion of trophoblast cells through regulation of Akt signaling. Reprod Toxicol 2013; 39:33-9. [PMID: 23571125 DOI: 10.1016/j.reprotox.2013.03.011] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/05/2012] [Revised: 03/20/2013] [Accepted: 03/28/2013] [Indexed: 01/07/2023]
Abstract
Recently, Viscum album var. coloratum agglutinin (VCA) was shown to have various effects on cancer cells. However, most researchers are focused on high concentrations (1-1000 ng/ml) of VCA and its anti-cancer effects. Therefore, we wanted to know whether low concentrations of VCA have an effect on proliferation and invasion of human trophoblast cells (HTR-8/SVneo cell line). Cell proliferations at low concentration of VCA (1-10 pg/ml) were increased over 2-fold relative to the control. Also, gelatinolytic activities of matrix metalloproteinase-2 were increased after VCA treatment, while TIMP-1 expression was reduced. Furthermore, the expression of integrin subunits α5 and β1 were increased 1.5-fold when cells were treated with low dose of VCA (10 pg/ml). Lastly, VCA was able to promote trophoblast invasion through activation of the Akt signaling pathway. In conclusion, low concentrations of VCA can stimulate the ability of trophoblast cells to invade through the extracellular matrix in vitro.
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Affiliation(s)
- Su-Yun Lyu
- College of Pharmacy, Sunchon National University, Megok-Dong, Suncheon-Si, Jeonnam 540-742, Republic of Korea
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Choi JH, Lyu SY, Lee HJ, Jung J, Park WB, Kim GJ. Korean mistletoe lectin regulates self-renewal of placenta-derived mesenchymal stem cells via autophagic mechanisms. Cell Prolif 2012; 45:420-9. [PMID: 22925501 DOI: 10.1111/j.1365-2184.2012.00839.x] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/10/2023] Open
Abstract
OBJECTIVES The balance between survival and death is a key point for regulation of physiology of stem cells. Recently, applications of natural products to enhance efficiencies in culturing and differentiation of stem cells are increasing. Korean mistletoe lectin (Viscum album L. var. coloratum agglutinin, VCA) has been known to be toxic to some cancer cells, but it is still unclear whether VCA has a cytotoxic or indeed a proliferative effect on mesenchymal stem cells (MSCs). Here, we have compared effects of VCA in naïve placenta-derived stem cells (PDSCs), immortalized PDSCs and cancer cells (HepG2), and analysed their mechanisms. MATERIALS AND METHODS MTT assay was performed to analyse effects of VCA on naïve PDSCs, immortalized PDSCs and HepG2. FACS, ROS, caspase-3 assay, western blotting and immunofluorescence were performed to detect signalling events involved in self-renewal of the above cell types. RESULTS VCA had cancer cell-specific toxicity to HepG2 cells even with low concentrations of VCA (1-5 pg/ml), toxicity was observed to immortalized PDSCs and HepG2s, while proliferation of naïve PDSCs was significantly increased (P < 0.05). ROS production by VCA treatment in naïve PDSCs was significantly lower compared to controls (P < 0.05). Furthermore, autophagy was activated in naïve PDSCs treated with VCA through increase in type II LC3 and decrease in phosphorylated mTOR. CONCLUSIONS VCA can promote MSC proliferation through an activated autophagic mechanism.
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Affiliation(s)
- J H Choi
- Department of Biomedical Science, CHA University, Kangnak-ku, Seoul, South Korea
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Fermented mistletoe extract as a multimodal antitumoral agent in gliomas. EVIDENCE-BASED COMPLEMENTARY AND ALTERNATIVE MEDICINE 2012; 2012:501796. [PMID: 23133496 PMCID: PMC3485514 DOI: 10.1155/2012/501796] [Citation(s) in RCA: 39] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 07/26/2012] [Accepted: 09/05/2012] [Indexed: 01/03/2023]
Abstract
In Europe, commercially available extracts from the white-berry mistletoe (Viscum album L.) are widely used as a complementary cancer therapy. Mistletoe lectins have been identified as main active components and exhibit cytotoxic effects as well as immunomodulatory activity. Since it is still not elucidated in detail how mistle toe extracts such as ISCADOR communicate their effects, we analyzed the mechanisms that might be responsible for their antitumoral function on a molecular and functional level. ISCADOR-treated glioblastoma (GBM) cells down-regulate central genes involved in glioblastoma progression and malignancy such as the cytokine TGF-β and matrix-metalloproteinases. Using in vitro glioblastoma/immune cell co-cultivation assays as well as measurement of cell migration and invasion, we could demonstrate that in glioblastoma cells, lectin-rich ISCADOR M and ISCADOR Q significantly enforce NK-cell-mediated GBM cell lysis. Beside its immune stimulatory effect, ISCADOR reduces the migratory and invasive potential of glioblastoma cells. In a syngeneic as well as in a xenograft glioblastoma mouse model, both pretreatment of tumor cells and intratumoral therapy of subcutaneously growing glioblastoma cells with ISCADOR Q showed delayed tumor growth. In conclusion, ISCADOR Q, showing multiple positive effects in the treatment of glioblastoma, may be a candidate for concomitant treatment of this cancer.
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Park YK, Do YR, Jang BC. Apoptosis of K562 leukemia cells by Abnobaviscum F®, a European mistletoe extract. Oncol Rep 2012; 28:2227-32. [PMID: 22972372 DOI: 10.3892/or.2012.2026] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/19/2012] [Accepted: 08/06/2012] [Indexed: 11/06/2022] Open
Abstract
Evidence suggests that mistletoe extract has the potential to be used as an anticancer agent. Abnobaviscum F® is a European mistletoe extract from the host tree Fraxinus. We investigated the effect of Abnobaviscum F on the growth and survival of different leukemia cell lines. Abnobaviscum F treatment strongly reduced survival and induced apoptosis of K562 (human myeloid leukemia), RPMI-8226 (human plasmacytoma) and L1210 (murine lymphocytic leukemia) cells in culture. Using K562 cells to further investigate the mechanism of action of Abnobaviscum F, we showed that Abnobaviscum F-induced cell death was associated with the activation of caspase-9, JNK-1/2 and p38 MAPK, as well as with the downregulation of Mcl-1, and inhibition of ERK-1/2 and PKB phosphorylation. Moreover, Abnobaviscum F treatment led to both a reduction of cellular glutathione (GSH) and the induction of ER stress (GRP78 and CHOP induction and eIF-2α phosphorylation). By contrast, Abnobaviscum F did not impact the expression of the DR4 and DR5 death receptors. The Abnobaviscum F-induced apoptosis of K562 cells was blocked by pretreatment with either GSH, z-VAD-fmk or SP600125. Our results, therefore, show that Abnobaviscum F induces apoptosis of K562 cells through the activation of the intrinsic caspase pathway, the phosphorylation of JNK-1, the reduction of cellular GSH, and the induction of ER stress.
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Affiliation(s)
- Yu-Kyoung Park
- Department of Medical Genetic Engineering, School of Medicine, Keimyung University, Dalseo-Gu, Daegu 704-701, Republic of Korea
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Faheina-Martins GV, da Silveira AL, Cavalcanti BC, Ramos MV, Moraes MO, Pessoa C, Araújo DAM. Antiproliferative effects of lectins from Canavalia ensiformis and Canavalia brasiliensis in human leukemia cell lines. Toxicol In Vitro 2012; 26:1161-9. [PMID: 22776218 DOI: 10.1016/j.tiv.2012.06.017] [Citation(s) in RCA: 41] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/15/2012] [Revised: 06/16/2012] [Accepted: 06/29/2012] [Indexed: 01/19/2023]
Abstract
The antiproliferative activity of lectins Canavalia ensiformis (ConA) and Canavalia brasiliensis (ConBr) were studied using human leukemia MOLT-4 and HL-60 cell lines. It was revealed that both ConA and ConBr were markedly cytotoxic to cells using MTT and NAC assays. The IC(50) values were approximately 3 and 20 μg/mL for ConA and ConBr, respectively, for both MOLT-4 and HL-60 cells. However, in normal human peripheral blood lymphocytes, the lectins were not cytotoxic, even when tested at concentrations as high as 200 μg/ml. Using comet assay, the lectins produced a rate of DNA damage exceeding 80% in MOLT-4 and HL-60 cells. Fluorescence analysis revealed the morphology characteristic of apoptosis, with low concentrations of apoptotic bodies and fragmented DNA (5 μg/ml). Flow cytometric analysis demonstrated an accumulation of cells in the sub-G1 cell cycle that is characteristic of DNA fragmentation, and a decrease in membrane integrity at high concentrations. Lastly, we evaluated the alterations in mitochondrial potential that reduced after treatment with lectins. Our results indicate that ConA and ConBr inhibited cell proliferation selectively in tumor cells and that apoptosis was the main death mechanism. Therefore, lectins can be considered a class of molecules with a high antitumor activity potential.
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Affiliation(s)
- Glaucia V Faheina-Martins
- Departamento de Biotecnologia, Centro de Biotecnologia, Universidade Federal da Paraíba, João Pessoa, PB, Brazil
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Delebinski CI, Jaeger S, Kemnitz-Hassanin K, Henze G, Lode HN, Seifert GJ. A new development of triterpene acid-containing extracts from Viscum album L. displays synergistic induction of apoptosis in acute lymphoblastic leukaemia. Cell Prolif 2012; 45:176-87. [PMID: 22221251 DOI: 10.1111/j.1365-2184.2011.00801.x] [Citation(s) in RCA: 27] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/23/2022] Open
Abstract
OBJECTIVES Aqueous Viscum album L. extracts are widely used for anti-cancer therapies. Due to their low solubility, triterpenes (which are known to act on cancers), do not occur in aqueous extracts in significant amounts. Using cyclodextrins, we have found it possible to solubilize mistletoe triterpene acids and to determine their effects on acute lymphoblastic leukaemia (ALL) in vitro and in vivo. MATERIALS AND METHODS A C.B-17/SCID model of pre-B ALL (NALM-6) was used to test efficacy and mechanisms of treatment with lectin- and triterpene acid containing preparations in vivo. Cytotoxicity of increasing concentrations of V. album L. preparations was assessed in vitro. Apoptosis was determined using mitochondrial membrane potential measurements, annexin V/PI, western blot analyses and caspase inhibitor assays. RESULTS Solubilized triterpene acid- or lectin-containing V. album L. extracts inhibited cell proliferation and demonstrated cytotoxic properties in vitro. Annexin V/PI and mitochondrial membrane potential assays indicated that dose-dependent induction of apoptosis was the main mechanism. Combination (viscumTT) of lectin- (viscum) and triterpene-containing (TT) extracts resulted in greatest induction of apoptosis. Furthermore, caspase activity demonstrated that these extracts were able to induce apoptosis through both caspase-8 and -9 dependent pathways. In vivo experimentation showed that treatment of mice with viscumTT combination prolonged mean survival to 50.5 days compared to 39.3 days in the phosphate-buffered saline group. CONCLUSION Here for the first time, we have demonstrated that either solubilized triterpene acids or lectins and combinations thereof, induce dose-dependent apoptosis in the ALL cell line NALM-6 via caspase-8 and -9 dependent pathways.
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Affiliation(s)
- C I Delebinski
- Department of Paediatrics, Division of Oncology/Haematology, Otto Heubner Centre for Paediatric and Adolescent Medicine (OHC), Charité, Universitaetsmedizin Berlin, Germany
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15
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García-Gasca T, García-Cruz M, Hernandez-Rivera E, López-Matínez J, Castañeda-Cuevas AL, Yllescas-Gasca L, Rodríguez-Méndez AJ, Mendiola-Olaya E, Castro-Guillén JL, Blanco-Labra A. Effects of Tepary bean (Phaseolus acutifolius) protease inhibitor and semipure lectin fractions on cancer cells. Nutr Cancer 2012; 64:1269-78. [PMID: 23163855 PMCID: PMC3856472 DOI: 10.1080/01635581.2012.722246] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/08/2012] [Accepted: 07/29/2012] [Indexed: 11/12/2022]
Abstract
Some natural and synthetic protease inhibitors (PI), such as the Bowman-Birk PI from soybean, have anticancer effects. We previously purified and characterized a Bowman-Birk-type PI from Tepary bean (Phaseolus acutifolius) seeds (TBPI). A semipure protein fraction containing this inhibitor, when tested its in vitro effect on transformed cells, showed a differential cytotoxic effect, as well as an increase in cell attachment to culture dishes. In this article we report that lectins were responsible for the cytotoxic effect previously observed, exhibiting a differential, antiproliferative effect on nontransformed cells and on different lineages of cancer cells. Although the purified TBPI lacked cytotoxicity, it was found to be responsible for the increase in cell adhesion, decreasing culture dishes' extracellular matrix degradation, leading to a decrease of the in vitro cell invasion capacity. This effect coincided with the suppression of Matrix Metalloproteinase-9 activity. These results indicate that Tepary bean seeds contain at least 2 different groups of bioactive proteins with distinct effects on cancer cells.
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Affiliation(s)
- Teresa García-Gasca
- Facultad de Ciencias Naturales, Universidad Autónoma de Querétaro, Querétaro, México.
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16
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Hong CE, Lyu SY. The Antimutagenic Effect of Mistletoe Lectin (Viscum album
L. var. coloratum
agglutinin). Phytother Res 2011; 26:787-90. [DOI: 10.1002/ptr.3639] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/02/2011] [Revised: 06/27/2011] [Accepted: 07/11/2011] [Indexed: 11/06/2022]
Affiliation(s)
- Chang-Eui Hong
- Department of Pharmacy; Yeungnam University; 214-1 Dae-dong Gyeongsan Gyeongbuk 712-749 South Korea
| | - Su-Yun Lyu
- Department of Pharmacy; Sunchon National University; 255 Jungangno Suncheon Jeonnam 540-742 South Korea
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17
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Lyu SY, Park WB. Gene network analysis on the effect of Viscum album var. coloratum in T cells stimulated with anti-CD3/CD28 antibodies. Arch Pharm Res 2011; 34:1735-49. [DOI: 10.1007/s12272-011-1018-y] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/31/2010] [Revised: 04/27/2011] [Accepted: 06/08/2011] [Indexed: 01/15/2023]
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18
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Wang TH, Kung YL, Lee MH, Su NW. N-acetyl-D-galactosamine-specific lectin isolated from the seeds of Carica papaya. JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 2011; 59:4217-4224. [PMID: 21405109 DOI: 10.1021/jf104962g] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/30/2023]
Abstract
N-Acetyl-D-galactosamine (GalNAc)-specific lectins are of great interest because they have been reported to detect tumor-associated antigens of malignant cells. We isolated a novel lectin from Carica papaya seeds, named C. papaya lectin (CPL). Purification of the lectin involved ammonium sulfate fractionation and DEAE anion exchange and repeated gel filtration chromatography. Inhibition of CPL causing hemagglutination on human erythrocytes showed that the lectin shows specificity to GalNAc and lactose. Surface plasmon resonance further revealed that the lectin possesses high specificity toward GalNAc with a dissociation constant of 5.5 × 10(-9) M. The lectin is composed of 38- and 40-kDa subunits with a molecular mass of ∼804 kDa estimated by size-exclusion high-performance liquid chromatography. Incubation of CPL with Jurkat T cells showed significant induction of IL-2 cytokine, which suggests that CPL has potent immunomodulatory effects on immune cells.
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Affiliation(s)
- Teng-Hsu Wang
- Department of Agricultural Chemistry, National Taiwan University, Taipei, Taiwan
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19
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Lyu SY, Park WB. Effect of Korean Mistletoe Lectin on Gene Expression Profile in Human T Lymphocytes: A Microarray Study. Biomol Ther (Seoul) 2010. [DOI: 10.4062/biomolther.2010.18.4.411] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/05/2022] Open
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20
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Yan Q, Li Y, Jiang Z, Sun Y, Zhu L, Ding Z. Antiproliferation and apoptosis of human tumor cell lines by a lectin (AMML) of Astragalus mongholicus. PHYTOMEDICINE : INTERNATIONAL JOURNAL OF PHYTOTHERAPY AND PHYTOPHARMACOLOGY 2009; 16:586-593. [PMID: 19403285 DOI: 10.1016/j.phymed.2008.12.024] [Citation(s) in RCA: 44] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 11/11/2008] [Accepted: 12/19/2008] [Indexed: 05/27/2023]
Abstract
A lectin (AMML) from the roots of Astragalus mongholicus was extracted and purified by affinity chromatographic technique. Human cervical carcinoma cell line (HeLa), human osteoblast-like cell line (MG63) and human leukemia cell line (K562) were used to check the effects of AMML on cell proliferation, apoptosis and cell cycle. Maximum growth inhibition (92%) was observed with HeLa cells, followed by K562 cells (84%) and MG63 (48%) cells. Morphological observation showed that AMML-treated HeLa cells displayed outstanding apoptosis characteristics, such as nuclear fragmentation and appearance of membrane-enclosed apoptotic bodies. The apoptosis of HeLa cells was confirmed by flow cytometry using Annexin V/FITC and propidium iodide (PI) staining technique. For the first time we also report a significant cell cycle arrest at S phase of HeLa cells by AMML. Therefore, the present investigation may lead to the possible therapeutic use of Astragalus mongholicus lectin.
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Affiliation(s)
- Qiaojuan Yan
- Bioresource Utilization Laboratory, College of Engineering, China Agricultural University, Beijing 100083, China
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21
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Lyu SY, Park WB. Mistletoe lectin modulates intestinal epithelial cell-derived cytokines and B cell IgA secretion. Arch Pharm Res 2009; 32:443-51. [DOI: 10.1007/s12272-009-1319-6] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/10/2008] [Revised: 01/19/2009] [Accepted: 02/26/2009] [Indexed: 12/15/2022]
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22
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23
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Transport of mistletoe lectin by M cells in human intestinal follicle-associated epithelium (FAE) In vitro. Arch Pharm Res 2008; 31:1613-21. [DOI: 10.1007/s12272-001-2159-1] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/05/2008] [Revised: 12/05/2008] [Accepted: 12/06/2008] [Indexed: 11/24/2022]
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24
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Seifert G, Jesse P, Laengler A, Reindl T, Lüth M, Lobitz S, Henze G, Prokop A, Lode HN. Molecular mechanisms of mistletoe plant extract-induced apoptosis in acute lymphoblastic leukemia in vivo and in vitro. Cancer Lett 2008; 264:218-28. [PMID: 18314258 DOI: 10.1016/j.canlet.2008.01.036] [Citation(s) in RCA: 70] [Impact Index Per Article: 4.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/12/2007] [Revised: 01/16/2008] [Accepted: 01/18/2008] [Indexed: 11/19/2022]
Abstract
Viscum album (Mistletoe) is one of the most widely used alternative cancer therapies. Aqueous mistletoe extracts (MT) contain the three mistletoe lectins I, II and III as one predominant group of biologically active agents. Although MT is widely used, there is a lack of scientifically sound preclinical and clinical data. In this paper, we describe for the first time the in vivo efficacy and mechanism of action of MT in lymphoblastic leukemia. For this purpose, we first investigated both the cytotoxic effect and the mechanism of action of two standardized aqueous MTs (MT obtained from fir trees (MT-A); MT obtained from pine trees (MT-P)) in a human acute lymphoblastic leukemia (ALL) cell line (NALM-6). MT-A, MT-P and ML-I inhibited cell proliferation as determined by Casy Count analysis at very low concentrations with MT-P being the most cytotoxic extract. DNA-fragmentation assays indicated that dose-dependent induction of apoptosis was the main mechanism of cell death. Finally, we evaluated the efficacy of MT-A and MT-P in an in vivo SCID-model of pre-B ALL (NALM-6). Both MTs significantly improved survival (up to 55.4 days) at all tested concentrations in contrast to controls (34.6 days) without side effects.
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Affiliation(s)
- Georg Seifert
- Department of Pediatric Oncology/Hematology, Otto-Heubner-Center for Pediatric and Adolescent Medicine, Charité, Universitätsmedizin Berlin, Germany.
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25
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Effects of Korean Mistletoe Lectin(Viscum album coloratum) on Proliferation and Cytokine Expression in Human Peripheral Blood Mononuclear Cells and T-Lymphocytes. Arch Pharm Res 2007; 30:1252-64. [DOI: 10.1007/bf02980266] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/21/2022]
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26
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Khil LY, Kim W, Lyu S, Park WB, Yoon JW, Jun HS. Mechanisms involved in Korean mistletoe lectin-induced apoptosis of cancer cells. World J Gastroenterol 2007; 13:2811-8. [PMID: 17569116 PMCID: PMC4395632 DOI: 10.3748/wjg.v13.i20.2811] [Citation(s) in RCA: 44] [Impact Index Per Article: 2.4] [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 investigate the anti-cancer mechanisms of Korean mistletoe lectin (Viscum album coloratum agglutinin, VCA) using a human colon cancer cell line (COLO).
METHODS: Cytotoxic effects of VCA on COLO cells were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in vitro and tumor-killing effects in vivo. To study the mechanisms involved, the expression of various pro-caspases, anti-apoptotic proteins, and death receptors was determined by western blot. To determine which death receptor is involved in VCA-induced apoptosis of COLO cells, cytotoxicity was examined by MTT assay after treatment with agonists or antagonists of death receptors.
RESULTS: VCA killed COLO cells in a time- and dose-dependent manner and induced complete regression of tumors in nude mice transplanted with COLO cells. Treatment of COLO cells with VCA activated caspase-2, -3, -8, and -9 and decreased expression of anti-apoptotic molecules including receptor interacting protein, nuclear factor-κB, X-linked inhibitor of apoptosis protein, and Akt/protein kinase B. We then examined the involvement of death receptors in VCA-induced apoptosis. Only tumor necrosis factor receptor 1, among the death receptors examined, was involved in apoptosis of COLO cells, evidenced by inhibition of VCA-induced apoptosis and decreased activation of caspases, particularly caspase-8, by tumor necrosis factor receptor 1 antagonizing antibody.
CONCLUSION: VCA-induced apoptotic COLO cell death is due to the activation of caspases and inhibition of anti-apoptotic proteins, in part through the tumor necrosis factor receptor 1 signaling pathway.
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Affiliation(s)
- Lee-Yong Khil
- Department of Microbiology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada
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27
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Lyu SY, Park WB. Mistletoe lectin (Viscum album coloratum) modulates proliferation and cytokine expressions in murine splenocytes. BMB Rep 2007; 39:662-70. [PMID: 17129400 DOI: 10.5483/bmbrep.2006.39.6.662] [Citation(s) in RCA: 15] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
It is well documented that an extract of European mistletoe has a variety of biological effects, such as the stimulation of cytokine production from immune cells, and additional immunoadjuvant activities. While the European mistletoe has been studied intensively, we know less about Korean mistletoe as a therapeutic plant, especially as a possible immunomodulating drug. This study will investigated the effects of Korean mistletoe lectin (Viscum album L. var. coloratum agglutinin, VCA) on murine splenocytes to investigate whether VCA acts as an immunomodulator, which could lead to improved immune responses in these cells. The results showed that VCA inhibited cell proliferation at higher concentrations (at 1-8 ng/ml) and enhanced cell proliferation at lower concentrations (at 4-32 pg/ml). Further studies were carried out to determine if the proproliferative or anti-proliferative activity exhibited by VCA was correlated with cytokine secretion. Consequently, interferon (IFN)-gamma secretion was decreased in concanavalin A (ConA)-stimulated murine splenocytes by VCA (4-64 ng/ml), but there was no change in IL-4 levels. This suggests that VCA has the ability to modulate murine splenocyte proliferation and can possibly act on the balance of Th1/Th2 cellular immune responses.
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Affiliation(s)
- Su-Yun Lyu
- Immune Modulation Research Group, School of Pharmacy, University of Nottingham, University Park, Nottingham, NG7 2RD, United Kingdom.
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28
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Li X, Feng P, Wen ZY, Zhu M, Liu SH, Wang XJ. IGrowth inhibition of human hepatocarcinoma cells induced by Chinese herbs Huqi San and its principal drug mistletoe extracts. Shijie Huaren Xiaohua Zazhi 2006; 14:1963-1969. [DOI: 10.11569/wcjd.v14.i20.1963] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: To probe the effect of Chinese herbs, Huqi San (HQS) and its principal drug mistletoe extracts on the growth of human hepatocar-cinoma cell line SMMC-7721 and the levels of intracellular reactive oxygen species (ROS).
METHODS: The proliferation of SMMC-7721 cells inhibited by HQS (0.3125, 0.625, 1.25 g/L), mistletoe polysaccharides (0.625, 1.25, 2.5, 5 g/L) and total alkaloid of mistletoe (3, 6, 12 g/L) at 48, 72 and 96 h were observed by MTT assay. The cell cycle and apoptosis rate of SMMC-7721 cells were examined by flow cytometry. The fluorescent intensity of ROS was observed by laser scanning confocal microscopy.
RESULTS: Both HQS and mistletoe extracts inhibited the proliferation of SMMC-7721 cells in a time- and dose-dependence manner, and the numeric value of absorbance in high-dose group decreased markedly as compared with that in the control group (HQS 72 h: 1.022 ± 0.13 vs 1.207 ± 0.04, P < 0.01; 96 h: 1.235 ± 0.20 vs 1.602 ± 0.05, P < 0.01; mistletoe polysaccharides 48 h: 0.570 ± 0.03 vs 0.744 ± 0.01, P < 0.01; 72 h: 0.803 ± 0.04 vs 1.207 ± 0.04, P < 0.01; 96 h: 0.860 ± 0.13 vs 1.602 ± 0.05, P < 0.01; total alkaloid of mistletoe 72 h: 0.919 ± 0.14 vs 1.233 ± 0.04, P < 0.01; 96 h: 0.701 ± 0.07 vs 1.819 ± 0.04, P < 0.01), expect that at 48 h in HQS and total alkaloid of mistletoe group. After treatment with the drugs for 72 h, the cell populations in the group of mistletoe polysaccharides and total alkaloid of mistletoe increased in G1 phase (81.0%, 86.9% vs 70.0%, P < 0.01) and decreased in G2 and S phase (13.1%, 5.7% vs 16.4%, P < 0.01; 5.9%, 7.4% vs 13.5%, P < 0.01), in comparison with those in the controls. In the group of HQS, the proportion of S-phase cells decreased (1.5% vs 13.5%, P < 0.01) while that of G2-phase ones increased (28.2% vs 16.4%, P < 0.01). However, there was no obvious change in the numbers of G1-phase cells. In addition, all of the three drugs enhanced the apoptosis of SMMC-7721 cells (HQS: 14.8% vs 6.0%, P < 0.01; mistletoe polysaccharides: 11.7% vs 6.0%, P < 0.01; total alkaloid of mistletoe: 6.7% vs 6.0%, P < 0.05). Confocal microscopy showed that the fluorescent intensity declined suddenly and then maintained at a low level almost without fluctuating when the three kinds of drugs were added. However, after the fluorescent intensity was stabile at a baseline, it increased instantly as the total alkaloid of mistletoe was added. Thereafter, it decreased rapidly and maintained at a low level for a period of time. But this phenomenon was not observed when the cells were treated with HQS and mistletoe polysaccharides.
CONCLUSION: HQS, mistletoe polysaccharides and total alkaloid of mistletoe can inhibit the proliferation and promote the apoptosis of SMMC-7721 cells.
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29
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Lyu SY, Moon YS, Park WB. Tyrosinase Inhibitory and Antioxidant Activities of Korean Mistletoe (Viscum album var. coloratum) Extract and Its Fractions. Prev Nutr Food Sci 2005. [DOI: 10.3746/jfn.2005.10.3.244] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022] Open
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30
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Kim WH, Park WB, Gao B, Jung MH. Critical role of reactive oxygen species and mitochondrial membrane potential in Korean mistletoe lectin-induced apoptosis in human hepatocarcinoma cells. Mol Pharmacol 2004; 66:1383-96. [PMID: 15340045 DOI: 10.1124/mol.104.001347] [Citation(s) in RCA: 75] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/08/2023] Open
Abstract
Viscum album L. coloratum agglutinin (VCA), isolated from Korean mistletoe, is a strong inducer of apoptosis in a variety of tumor cells; however, the underlying molecular mechanisms responsible are not clear. Here, we show that VCA induces apoptotic killing, as demonstrated by DNA fragmentation, Hoechst 33258 staining, terminal deoxynucleotidyl transferase dUTP nick-end labeling assay, and flow cytometry analysis in hepatocarcinoma Hep3B cells. VCA treatment results in a significant increase in reactive oxygen species (ROS) and loss of mitochondrial membrane potential (DeltaPsim). Furthermore, treatment with the antioxidant N-acetyl-L-cysteine reduces ROS induction by VCA, preventing apoptosis in Hep3B cells, indicating that oxidative stress is involved in VCA-mediated cell death. Our results also show rapid changes in mitochondrial transition permeability, Bax translocation, cytochrome c release, caspase-3 activity, and poly(ADP-ribose) polymerase degradation in Hep3B cells occurring in VCA-induced apoptosis. There is much evidence that implicates c-Jun NH2-terminal kinase (JNK) activation with apoptosis in a variety of cellular and animal models. In this study, we show that VCA induces JNK phosphorylation, which is abolished with pretreatment with a JNK inhibitor. Moreover, Hep3B cells overexpressing JNK1 or stress-activated protein kinase kinase (SEK1) seem to be more susceptible to cell death from ROS and loss of DeltaPsim induced by VCA, whereas expression of dominant-negative JNK1 or SEK1 in Hep3B cells do not. These data suggest that JNK phosphorylation may be a major regulator involved in VCA-induced apoptosis. Together, these results suggest that VCA induces apoptosis by inducing ROS production and a loss of DeltaPsim, in which JNK phosphorylation plays a critical role in these events.
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Affiliation(s)
- Won-Ho Kim
- Division of Metabolic Disease, Department of Biomedical Science, National Institutes of Health, Seoul, South Korea
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31
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Choi SH, Lyu SY, Park WB. Mistletoe lectin induces apoptosis and telomerase inhibition in human A253 cancer cells through dephosphorylation of Akt. Arch Pharm Res 2004; 27:68-76. [PMID: 14969342 DOI: 10.1007/bf02980049] [Citation(s) in RCA: 64] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/12/2023]
Abstract
Mistletoe lectin has been reported to induce apoptosis in different cancer cell lines in vitro and to show antitumor activity against a variety of tumors in animal models. We previously demonstrated the Korean mistletoe lectin (Viscum album var. coloratum, VCA)-induced apoptosis by down-regulation of Bcl-2 and telomerase activity and by up-regulation of Bax through p53- and p21-independent pathway in hepatoma cells. In the present study, we observed the induction of apoptotic cell death through activation of caspase-3 and the inhibition of telomerase activity through transcriptional down-regulation of hTERT in the VCA-treated A253 cells. We also observed the inhibition of telomerase activity and induction of apoptosis resulted from dephosphorylation of Akt in the survival signaling pathways. In addition, combining VCA with the inhibitors of phosphatidylinositol 3-kinase (PI3-kinase) upstream of Akt, wortmannin and LY294002 showed an additive inhibitory effect of telomerase activity. In contrast, the inhibitor of protein phosphatase 2A (PP2A), okadaic acid inhibited VCA-induced dephosphorylation of Akt and inhibition of telomerase activity. Taken together, VCA induces apoptotic cell death through Akt signaling pathway in correlated with the inhibition of telomerase activity and the activation of caspase-3. From these results, together with our previous studies, we suggest that VCA triggers molecular changes that resulting in the inhibition of cell growth and the induction of apoptotic cell death of cancer cells, which suggest that VCA may be useful as chemotherapeutic agent for cancer cells.
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Affiliation(s)
- Sang Ho Choi
- Brain Disease Research Center, School of Medicine, Ajou Univerisity, Suwon 442-749, Korea
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32
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Lau CBS, Ho CY, Kim CF, Leung KN, Fung KP, Tse TF, Chan HHL, Chow MSS. Cytotoxic activities of Coriolus versicolor (Yunzhi) extract on human leukemia and lymphoma cells by induction of apoptosis. Life Sci 2004; 75:797-808. [PMID: 15183073 DOI: 10.1016/j.lfs.2004.04.001] [Citation(s) in RCA: 107] [Impact Index Per Article: 5.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/09/2003] [Accepted: 04/02/2004] [Indexed: 01/02/2023]
Abstract
Coriolus versicolor (CV), also known as Yunzhi, is one of the commonly used Chinese medicinal herbs. Although recent studies have demonstrated its antitumour activities on cancer cells in vitro and in vivo, the exact mechanism is not fully elucidated. Hence, the objective of this study was to examine the in vitro cytotoxic activities of a standardized aqueous ethanol extract prepared from Coriolus versicolor on a B-cell lymphoma (Raji) and two human promyelocytic leukemia (HL-60, NB-4) cell lines using a MTT cytotoxicity assay, and to test whether the mechanism involves induction of apoptosis. Cell death ELISA was employed to quantify the nucleosome production resulting from nuclear DNA fragmentation during apoptosis. The present results demonstrated that CV extract at 50 to 800 microg/ml dose-dependently suppressed the proliferation of Raji, NB-4, and HL-60 cells by more than 90% (p < 0.01), with ascending order of IC50 values: HL-60 (147.3 +/- 15.2 microg/ml), Raji (253.8 +/- 60.7 microg/ml) and NB-4 (269.3 +/- 12.4 microg/ml). The extract however did not exert any significant cytotoxic effect on normal liver cell line WRL (IC50 > 800 microg/ml) when compared with a chemotherapeutic anticancer drug, mitomycin C (MMC), confirming the tumour-selective cytotoxicity. Nucleosome productions in HL-60, NB-4 and Raji cells were significantly increased by 3.6-, 3.6- and 5.6-fold respectively upon the treatment of CV extract, while no significant nucleosome production was detected in extract-treated WRL cells. The CV extract was found to selectively and dose-dependently inhibit the proliferation of lymphoma and leukemic cells possibly via an apoptosis-dependent pathway.
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MESH Headings
- Antineoplastic Agents/pharmacology
- Apoptosis/drug effects
- Cell Division/drug effects
- Cell Line, Tumor
- Cell Survival/drug effects
- DNA Fragmentation
- Dose-Response Relationship, Drug
- Drug Screening Assays, Antitumor
- Drugs, Chinese Herbal/pharmacology
- Formazans/metabolism
- Humans
- Leukemia, Promyelocytic, Acute/drug therapy
- Leukemia, Promyelocytic, Acute/metabolism
- Leukemia, Promyelocytic, Acute/pathology
- Liver/drug effects
- Liver/pathology
- Lymphoma, B-Cell/drug therapy
- Lymphoma, B-Cell/metabolism
- Lymphoma, B-Cell/pathology
- Medicine, Chinese Traditional
- Mitomycin/pharmacology
- Nucleosomes/drug effects
- Plants, Medicinal/chemistry
- Tetrazolium Salts/metabolism
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Affiliation(s)
- C B S Lau
- School of Pharmacy, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.
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Liu WK, Sze SCW, Ho JCK, Liu BPL, Yu MC. Wheat germ lectin induces G2/M arrest in mouse L929 fibroblasts. J Cell Biochem 2004; 91:1159-73. [PMID: 15048871 DOI: 10.1002/jcb.10755] [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] [Indexed: 11/08/2022]
Abstract
Wheat germ lectin (WGA) is a cytotoxic lectin for many cell lines [Wang et al., 2000], but its underlying mechanism is not clear. In this report, we found that incubation of synchronized mouse L929 fibroblasts with WGA resulted in a dose-dependent reduction of intracellular incorporation of 3H-thymidine and MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide)-conversion activity (IC50 congruent with 0.4 microM). Fluorescein-conjugated WGA was demonstrated to transport from the cell surface into the paranuclear region of cultured L929 cells within 30 min, and subsequently evoked lipid peroxidation of plasma membrane and vacuolation in the cytoplasm of these cells. Studies with tritiated thymidine incorporation, immunofluorescence microscopy, immunoblotting analysis and flow cytometry revealed that WGA inhibited cell cycle progression after one replication, resulting in G2/M arrest and alteration of cell cycle regulatory proteins, particularly activation of p21Cip1/WAF1 and suppression of cyclin B and cdc 2. Although there was an increase of cytosolic caspase 3 and bax protein expression, no apoptotic bodies were observed by both fluorescence and transmission electron microscopy. These results suggest that WGA arrested L929 proliferation after one cell cycle in the G2/M phase through activation of the p21Cip1/WAF1 and suppression of Cyclin B-Cdc2.
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Affiliation(s)
- W K Liu
- Department of Anatomy, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.
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Lyu SY, Kwon YJ, Joo HJ, Park WB. Preparation of alginate/chitosan microcapsules and enteric coated granules of mistletoe lectin. Arch Pharm Res 2004; 27:118-26. [PMID: 14969350 DOI: 10.1007/bf02980057] [Citation(s) in RCA: 18] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Abstract
The aqueous extract of European mistletoe (Viscum album, L.) has been used in cancer therapy. The purified mistletoe lectins, main components of mistletoe, have demonstrated cytotoxic and immune-system-stimulating activities. Korean mistletoe (Viscum album L. coloratum), a subspecies of European mistletoe, has also been reported to possess anticancer and immunological activities. A galactose- and N-acetyl-D-galactosamine-specific lectin (Viscum album L. coloratum agglutinin, VCA) with Mr 60 kDa was isolated from Korean mistletoe. Mistletoe preparations have been given subcutaneously due to the low stability of lectin in the gastrointestinal (GI) tract. In the present study, we investigated the possibility of alginate/chitosan microcapsules as a tool for oral delivery of mistletoe lectin. In addition, our strategy has been to develop a system composed of stabilizing cores (granules), which contain mistletoe lectin, extract or powder, coated by a biodegradable polymer wall. Our results indicated that successful incorporation of VCA into alginate/chitosan microcapsules has been achieved and that the alginate/chitosan microcapsule protected the VCA from degradation at acidic pH values. And coating the VCA with polyacrylic polymers, Eudragit, produced outstanding results with ideal release profiles and only minimal losses of cytotoxicity after manufacturing step. The granules prepared with extract or whole plant produced the best results due to the stability in the extract or whole plant during manufacturing process.
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Affiliation(s)
- Su-Yun Lyu
- College of Natural Sciences, Seoul Womens University, Seoul 139-774, Korea
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Abstract
The control of cancer, the second leading cause of death worldwide, may benefit from the potential that resides in alternative therapies. The primary carcinogens stem from a variety of agricultural, industrial, and dietary factors. Conventional therapies cause serious side effects and, at best, merely extend the patient's lifespan by a few years. There is thus the need to utilise alternative concepts or approaches to the prevention of cancer. This review focuses on the many natural products that have been implicated in cancer prevention and that promote human health without recognisable side effects. These molecules originate from vegetables, fruits, plant extracts, and herbs.
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Affiliation(s)
- L Reddy
- Department of Biotechnology, Durban Institute of Technology, P.O. Box 1334, Durban 4000, South Africa
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Van Huyen JPD, Bayry J, Delignat S, Gaston AT, Michel O, Bruneval P, Kazatchkine MD, Nicoletti A, Kaveri SV. Induction of Apoptosis of Endothelial Cells by Viscum album: A Role for Anti-Tumoral Properties of Mistletoe Lectins. Mol Med 2002. [DOI: 10.1007/bf03402170] [Citation(s) in RCA: 17] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/28/2022] Open
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Lyu SY, Choi SH, Park WB. Korean mistletoe lectin-induced apoptosis in hepatocarcinoma cells is associated with inhibition of telomerase via mitochondrial controlled pathway independent of p53. Arch Pharm Res 2002; 25:93-101. [PMID: 11885700 DOI: 10.1007/bf02975269] [Citation(s) in RCA: 81] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
Abstract
The extract of European mistletoe (Viscum album, L) has been used in adjuvant chemotherapy of cancer and mistletoe lectins are considered to be major active components. The present work was performed to investigate the effects of Korean mistletoe lectin (Viscum album L. coloratum agglutinin, VCA) on proliferation and apoptosis of human hepatoma cells as well as the underlying mechamisms for these effects. We showed that VCA induced apoptosis in both SK-Hep-1 (p53-positive) and Hep 3B (p53-negative) cells through p53- and p21-independent pathways. VCA induced apoptosis by down-regulation of Bcl-2 and by up-regulation of Bax functioning upstream of caspase-3 in both cell lines. In addition, we observed down-regulation of telomerase activity in both VCA-treated cells. Our results provide direct evidence of the anti-tumor potential of this biological response which comes from inhibition of telomerase and consequent inducing apoptosis. VCA-induced apoptosis is regulated by mitochondrial controlled pathway independently of p53. These findings are important for the therapy with preparation of mistletoe because they show that telomerase-dependent mechanism can be targeted by VCA in human hepatocarcinoma. Taken together, our results suggest that the VCA, considered as a telomerase-inhibitor, can be envisaged as a candidate for enhancing sensitivity of conventional anticancer drugs.
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MESH Headings
- Antineoplastic Agents, Phytogenic/pharmacology
- Apoptosis/drug effects
- Blotting, Western
- Carcinoma, Hepatocellular/enzymology
- Carcinoma, Hepatocellular/pathology
- Caspase 3
- Caspase Inhibitors
- Cell Nucleus/pathology
- Cell Nucleus/ultrastructure
- DNA Fragmentation/drug effects
- DNA, Neoplasm/drug effects
- Down-Regulation/drug effects
- Enzyme Inhibitors/pharmacology
- Flow Cytometry
- Genes, bcl-2/drug effects
- Genes, bcl-2/genetics
- Genes, p53/drug effects
- Humans
- Korea
- Mitochondria/enzymology
- Neoplasm Proteins/chemistry
- Neoplasm Proteins/isolation & purification
- Oncogene Protein p21(ras)/genetics
- Plant Preparations
- Plant Proteins
- Proto-Oncogene Proteins/genetics
- Proto-Oncogene Proteins c-bcl-2
- Ribosome Inactivating Proteins, Type 2
- Telomerase/antagonists & inhibitors
- Toxins, Biological/pharmacology
- Tumor Cells, Cultured
- bcl-2-Associated X Protein
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Affiliation(s)
- Su Yun Lyu
- College of Natural Sciences, Seoul Women's University, Seoul, Korea
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Park WB, Lyu SY, Kim JH, Choi SH, Chung HK, Ahn SH, Hong SY, Yoon TJ, Choi MJ. Inhibition of tumor growth and metastasis by Korean mistletoe lectin is associated with apoptosis and antiangiogenesis. Cancer Biother Radiopharm 2001; 16:439-47. [PMID: 11776761 DOI: 10.1089/108497801753354348] [Citation(s) in RCA: 47] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/12/2022] Open
Abstract
The mistletoe lectins are major active components in the extract of European mistletoes that have been widely used in adjuvant chemotherapy of cancer. This study was performed to investigate the mechanism of anticancer and antimetastatic activity of the purified Korean mistletoe lectin (Viscum album L. coloratum agglutinin, VCA). C57BL6 mice inoculated with B16-BL6 melanoma cells and treated with VCA were assessed for survival and metastasis. The induction of apoptosis of B16-BL6 cells by VCA was investigated by morphological changes, DNA fragmentation characteristics, and cell cycle analysis. The antiangiogenic activity of VCA was also measured by the CAM (choriallantoic membrane) assay. Length of survival of mice was increased and lung metastasis was inhibited by VCA. Treatment of cells with VCA resulted in growth suppression, nuclear morphological changes, DNA fragmentation, and an increased fraction of cells in sub-G1 consistent with apoptosis. Antiangiogenesis of VCA was assessed by CAM assay, where vessel growth induced by fat emulsion was decreased. These results suggest that VCA inhibits tumor growth and metastasis by increasing apoptosis and inhibiting angiogenesis.
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MESH Headings
- Allantois/blood supply
- Allantois/drug effects
- Angiogenesis Inhibitors/pharmacology
- Angiogenesis Inhibitors/therapeutic use
- Animals
- Antineoplastic Agents, Phytogenic/therapeutic use
- Apoptosis/drug effects
- Cell Cycle/drug effects
- Chick Embryo
- Chorion/blood supply
- Chorion/drug effects
- Drug Screening Assays, Antitumor
- Female
- Lung Neoplasms/drug therapy
- Lung Neoplasms/secondary
- Melanoma, Experimental/blood supply
- Melanoma, Experimental/drug therapy
- Melanoma, Experimental/pathology
- Melanoma, Experimental/secondary
- Mice
- Mice, Inbred C57BL
- Neovascularization, Pathologic/drug therapy
- Neovascularization, Physiologic/drug effects
- Plant Preparations
- Plant Proteins
- Ribosome Inactivating Proteins, Type 2
- Toxins, Biological/therapeutic use
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Affiliation(s)
- W B Park
- College of Natural Science, Seoul Women's University, Seoul 139-774, Korea.
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