DOI QR코드

DOI QR Code

Condurango (Gonolobus condurango) Extract Activates Fas Receptor and Depolarizes Mitochondrial Membrane Potential to Induce ROS-dependent Apoptosis in Cancer Cells in vitro -CE-treatment on HeLa: a ROS-dependent mechanism-

  • Bishayee, Kausik (Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani) ;
  • Mondal, Jesmin (Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani) ;
  • Sikdar, Sourav (Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani) ;
  • Khuda-Bukhsh, Anisur Rahman (Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani)
  • Received : 2015.04.23
  • Accepted : 2015.06.11
  • Published : 2015.09.30

Abstract

Objectives: Condurango (Gonolobus condurango) extract is used by complementary and alternative medicine (CAM) practitioners as a traditional medicine, including homeopathy, mainly for the treatment of syphilis. Condurango bark extract is also known to reduce tumor volume, but the underlying molecular mechanisms still remain unclear. Methods: Using a cervical cancer cell line (HeLa) as our model, the molecular events behind condurango extract's (CE's) anticancer effect were investigated by using flow cytometry, immunoblotting and reverse transcriptase-polymerase chain reaction (RT-PCR). Other included cell types were prostate cancer cells (PC3), transformed liver cells (WRL-68), and peripheral blood mononuclear cells (PBMCs). Results: Condurango extract (CE) was found to be cytotoxic against target cells, and this was significantly deactivated in the presence of N-acetyl cysteine (NAC), a scavenger of reactive oxygen species (ROS), suggesting that its action could be mediated through ROS generation. CE caused an increase in the HeLa cell population containing deoxyribonucleic acid (DNA) damage at the G zero/Growth 1 (G0/G1) stage. Further, CE increased the tumor necrosis factor alpha ($TNF-{\alpha}$) and the fas receptor (FasR) levels both at the ribonucleic acid (RNA) and the protein levels, indicating that CE might have a cytotoxic mechanism of action. CE also triggered a sharp decrease in the expression of nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$) both at the RNA and the protein levels, a possible route to attenuation of B-cell lymphoma 2 (Bcl-2), and caused an opening of the mitochondrial membrane's permeability transition (MPT) pores, thus enhancing caspase activities. Conclusion: Overall, our results suggest possible pathways for CE mediated cytotoxicity in model cancer cells.

Acknowledgement

Supported by : UGC

References

  1. Sudhakar A. History of cancer, ancient and modern treatment methods. J Cancer Sci Ther. 2009;1(2):1-4. https://doi.org/10.4172/1948-5956.1000001
  2. Hayden EC. Cutting off cancer's supply lines. Nature. 2009;458(7239):686-7. https://doi.org/10.1038/458686b
  3. Zhang G, Wang Y, Zhang Y, Wan X, Li J, Liu K, et al. Anti-cancer activities of tea epigallocatechin-3-gallate in breast cancer patients under radiotherapy. Curr Mol Med. 2012;12(2):163-76. https://doi.org/10.2174/156652412798889063
  4. Troselj KG, Kujundzic RN. Curcumin in combined cancer therapy. Curr Pharm Des. 2014;20(42):6682-96. https://doi.org/10.2174/1381612820666140826154601
  5. Yao S, To KK, Wang YZ, Yin C, Tang C, Chai S, et al. Polyoxypregnane steroids from the stems of Marsdenia tenacissima. J Nat Prod. 2014;77(9):2044-53. https://doi.org/10.1021/np500385b
  6. Ye B, Li J, Li Z, Yang J, Niu T, Wang S. Anti-tumor activity and relative mechanism of ethanolic extract of Marsdenia tenacissima (Asclepiadaceae) against human hematologic neoplasm in vitro and in vivo. J Ethnopharmacol. 2014;153(1):258-67. https://doi.org/10.1016/j.jep.2014.02.035
  7. Huang Z, Lin H, Wang Y, Cao Z, Lin W, Chen Q. Studies on the anti-angiogenic effect of Marsdenia tenacissima extract in vitro and in vivo. Oncol Lett. 2013;5(3):917-22. https://doi.org/10.3892/ol.2013.1105
  8. Bishayee K, Paul A, Ghosh S, Sikdar S, Mukherjee A, Biswas R, et al. Condurango-glycoside-A fraction of Gonolobus condurango induces DNA damage associated senescence and apoptosis via ROS-dependent p53 signalling pathway in HeLa cells. Mol Cell Biochem. 2013;382(1-2):173-83. https://doi.org/10.1007/s11010-013-1732-5
  9. Hayashi K, Wada K, Mitsuhashi H, Bando H, Takase M, Terada S, et al. Antitumor active glycosides from Condurango cortex. Chem Pharm Bull. 1980;28(6):1954-8. https://doi.org/10.1248/cpb.28.1954
  10. Sikdar S, Mukherjee A, Khuda-Bukhsh AR. Ethanolic extract of Marsdenia condurango ameliorates benzo[a] pyrene-induced lung cancer of rats: condurango ameliorates BaP-induced lung cancer in rats. J Pharmacopuncture. 2014;17(2):7-17. https://doi.org/10.3831/KPI.2014.17.011
  11. Chen J, Li X, Sun C, Pan Y, Schlunegger UP. Identification of polyoxypregnane glycosides from the stems of Marsdenia tenacissima by high-performance liquid chromatography/tandem mass spectrometry. Talanta. 2008;77(1):152-9. https://doi.org/10.1016/j.talanta.2008.05.054
  12. Ulmer AJ, Scholz W, Ernst M, Brandt E, Flad HD. Isolation and subfractionation of human peripheral blood mononuclear cells (PBMC) by density gradient centrifugation on percoll. Immunobiology. 1984;166(3):238-50. https://doi.org/10.1016/S0171-2985(84)80042-X
  13. Paul A, Das S, Das J, Samadder A, Bishayee K, Sadhukhan R, et al. Diarylheptanoid-myricanone isolated from ethanolic extract of Myrica cerifera shows anticancer effects on HeLa and PC3 cell lines: signalling pathway and drug-DNA interaction. J Integr Med. 2013;11(6):405-15. https://doi.org/10.3736/jintegrmed2013057
  14. Paul A, Bishayee K, Ghosh S, Mukherjee A, Sikdar S, Chakraborty D, et al. Chelidonine isolated from ethanolic extract of Chelidonium majus promotes apoptosis in HeLa cells through p38-p53 and PI3K/AKT signalling pathways. Zhong Xi Yi Jie He Xue Bao. 2012;10(9):1025-38.
  15. Mao Z, Ke Z, Gorbunova V, Seluanov A. Replicatively senescent cells are arrested in G1 and G2 phases. Aging. 2012;4(6):431-5. https://doi.org/10.18632/aging.100467
  16. Fulda S, Gorman AM, Hori O, Samali A. Cellular stress responses: cell survival and cell death. Int J Cell Biol. 2010;2010;ID214074:1-23.
  17. Hanahan D, Weinberg RA. The hallmarks of cancer. Cell. 2000;100(1):57-70. https://doi.org/10.1016/S0092-8674(00)81683-9
  18. Lee SH, Meng XW, Flatten KS, Loegering DA, Kaufmann SH. Phosphatidylserine exposure during apoptosis reflects bidirectional trafficking between plasma membrane and cytoplasm. Cell Death Differ. 2013;20(1):64-76. https://doi.org/10.1038/cdd.2012.93
  19. Rieger AM, Nelson KL, Konowalchuk JD, Barreda DR. Modified annexin V/propidium iodide apoptosis assay for accurate assessment of cell death. J Vis Exp. 2011;50:2597.
  20. Lichter P, Walczak H, Weitz S, Behrmann I, Krammer PH. The human APO-1 (APT) antigen maps to 10q23, a region that is syntenic with mouse chromosome 19. Genomics. 1992;14(1):179-80. https://doi.org/10.1016/S0888-7543(05)80302-7
  21. Klopfer A, Hasenjager A, Belka C, Schulze-Osthoff K, Dorken B, Daniel PT. Adenine deoxynucleotides fludarabine and cladribine induce apoptosis in a CD95/Fas receptor, FADD and caspase-8-independent manner by activation of the mitochondrial cell death pathway. Oncogene. 2004;23(58):9408-18. https://doi.org/10.1038/sj.onc.1207975
  22. Roy S, Nicholson DW. Cross-talk in cell death signaling. J Exp Med. 2000;192(8):21-6. https://doi.org/10.1084/jem.192.8.F21
  23. Papa S, Bubici C, Zazzeroni F, Franzoso G. Mechanisms of liver disease: crosstalk between the NF-$\kappa$B and JNK pathways. Biol Chem. 2009;390(10):965-76. https://doi.org/10.1515/BC.2009.111
  24. Fiandalo MV, Kyprianou N. Caspase control: protagonists of cancer cell apoptosis. Exp Oncol. 2012;34(3):165-75.
  25. Murphy MP. How mitochondria produce reactive oxygen species. Biochem J. 2009;417(1):1-13. https://doi.org/10.1042/BJ20081386
  26. Pucci B, Kasten M, Giordano A. Cell cycle and apoptosis. Neoplasia. 2000;2(4):291-9. https://doi.org/10.1038/sj.neo.7900101

Cited by

  1. Resveratrol analogue (E)-8-acetoxy-2-[2-(3,4-diacetoxyphenyl) ethenyl]-quinazoline induces apoptosis via Fas-mediated pathway in HL-60 human leukemia cells vol.36, pp.6, 2016, https://doi.org/10.3892/or.2016.5168
  2. An Idiopathic Thrombocytopenic Purpura Patient Treated With Homeopathy: A Case Report vol.1, pp.2, 2016, https://doi.org/10.20286/hpr-010271
  3. Punica granatum (pomegranate) leaves extract induces apoptosis through mitochondrial intrinsic pathway and inhibits migration and invasion in non-small cell lung cancer in vitro vol.80, 2016, https://doi.org/10.1016/j.biopha.2016.03.023