Bcl-X(L) is functionally non-equivalent for the regulation of growth and survival in human ovarian cancer cells.
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.


CONCLUSIONS
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.
Dodier
Phillipe
P
Département de Microbiologie et Infectiologie, Faculté de Médecine, Université de Sherbrooke, 3001, 12ième Avenue Nord, Sherbooke, Canada J1H 5N1.
Piché
Alain
A
D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D002448:Cell Adhesion / Q000502:physiology; D048708:Cell Growth Processes; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000502:physiology; D002945:Cisplatin / Q000494:pharmacology; D005260:Female; D006801:Humans; D010051:Ovarian Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D017239:Paclitaxel / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection; D051020:bcl-X Protein / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*
C494811:BCL2L1 protein, human; D034741:RNA, Small Interfering; D051020:bcl-X Protein; D017239:Paclitaxel; D002945:Cisplatin