Phillipe Dodier1, Alain Piché. 1. Département de Microbiologie et Infectiologie, Faculté de Médecine, Université de Sherbrooke, 3001, 12ième Avenue Nord, Sherbooke, Canada J1H 5N1.
Abstract
OBJECTIVE: To investigate the role of Bcl-X(L) on the growth and survival of human ovarian carcinoma cells. METHODS: In this study, we enforced down-regulation of Bcl-X(L) by RNA interference in ovarian carcinoma cell lines CaOV3, SKOV3ip1 and OVCAR3 cell lines expressing various levels of Bcl-X(L). We also established stable transfectants of OVCAR3 cells overexpressing Bcl-X(L). We recently showed that Bcl-2 regulates cell cycle progression in ovarian cancer cells. Thus, the effect of Bcl-X(L) modulation on the rate of cell growth was determined by XTT assay. To evaluate the role of Bcl-X(L) in drug-induced apoptosis, cisplatin- and paclitaxel-induced apoptosis were determined in vitro for each of the three cell lines. Ovarian tumor cells must acquire the ability to survive in non-adherent conditions to grow in ascetic fluids. To mimic loss of anchorage and investigate the role of Bcl-X(L) in this process, cells were cultured on Hydrogel-coated plates and nuclear fragmentation, caspase-3 activation and nuclear propidium iodide staining were used to determine apoptosis. RESULTS: We show that enforced down-regulation of Bcl-X(L) protein significantly affected the growth rates of CaOV3 whereas it had only minimal effect on the other two cell lines. Down-regulation of Bcl-X(L) enhanced the sensitivity of CaOV3, OVCAR3 and SKOV3ip1 to cisplatin and paclitaxel. The susceptibility to apoptosis induced by loss of anchorage was also increased but in a cell line-dependent manner. Overexpression of Bcl-X(L) slowed the growth of OVCAR3 cells and conferred resistance to drug-induced apoptosis and apoptosis induced by loss of anchorage. CONCLUSION: Altogether, these findings demonstrate that modulation of Bcl-X(L) provokes changes in ovarian cancer cell growth and survival that are cell line-specific. Consequently, therapeutic strategy for treatment of ovarian cancer that target Bcl-X(L) will likely yield variable responses.
OBJECTIVE: To investigate the role of Bcl-X(L) on the growth and survival of humanovarian carcinoma cells. METHODS: In this study, we enforced down-regulation of Bcl-X(L) by RNA interference in ovarian carcinoma cell lines CaOV3, SKOV3ip1 and OVCAR3 cell lines expressing various levels of Bcl-X(L). We also established stable transfectants of OVCAR3 cells overexpressing Bcl-X(L). We recently showed that Bcl-2 regulates cell cycle progression in ovarian cancer cells. Thus, the effect of Bcl-X(L) modulation on the rate of cell growth was determined by XTT assay. To evaluate the role of Bcl-X(L) in drug-induced apoptosis, cisplatin- and paclitaxel-induced apoptosis were determined in vitro for each of the three cell lines. Ovarian tumor cells must acquire the ability to survive in non-adherent conditions to grow in ascetic fluids. To mimic loss of anchorage and investigate the role of Bcl-X(L) in this process, cells were cultured on Hydrogel-coated plates and nuclear fragmentation, caspase-3 activation and nuclear propidium iodide staining were used to determine apoptosis. RESULTS: We show that enforced down-regulation of Bcl-X(L) protein significantly affected the growth rates of CaOV3 whereas it had only minimal effect on the other two cell lines. Down-regulation of Bcl-X(L) enhanced the sensitivity of CaOV3, OVCAR3 and SKOV3ip1 to cisplatin and paclitaxel. The susceptibility to apoptosis induced by loss of anchorage was also increased but in a cell line-dependent manner. Overexpression of Bcl-X(L) slowed the growth of OVCAR3 cells and conferred resistance to drug-induced apoptosis and apoptosis induced by loss of anchorage. CONCLUSION: Altogether, these findings demonstrate that modulation of Bcl-X(L) provokes changes in ovarian cancer cell growth and survival that are cell line-specific. Consequently, therapeutic strategy for treatment of ovarian cancer that target Bcl-X(L) will likely yield variable responses.
Authors: Elizabeth H Stover; Maria B Baco; Ofir Cohen; Yvonne Y Li; Elizabeth L Christie; Mukta Bagul; Amy Goodale; Yenarae Lee; Sasha Pantel; Matthew G Rees; Guo Wei; Adam G Presser; Maya K Gelbard; Weiqun Zhang; Ioannis K Zervantonakis; Patrick D Bhola; Jeremy Ryan; Jennifer L Guerriero; Joan Montero; Felice J Liang; Andrew D Cherniack; Federica Piccioni; Ursula A Matulonis; David D L Bowtell; Kristopher A Sarosiek; Anthony Letai; Levi A Garraway; Cory M Johannessen; Matthew Meyerson Journal: Mol Cancer Res Date: 2019-08-28 Impact factor: 5.852