Bcl-XL is functionally non-equivalent for the regulation of growth and survival in human ovarian cancer cells
Bcl-XL is functionally non-equivalent for the regulation of growth and survival in human ovarian cancer cells
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DOI：
10.1016/j.ygyno.2005.08.028
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发表时间：
2006-02-01
影响因子：
4.7
通讯作者：
Piché, A
中科院分区：
文献类型：
--
作者：
Dodier, P;Piché, A
Objective.探讨Bcl-X-L对人卵巢癌细胞生长和存活的影响。在本研究中，我们通过RNA干扰在表达不同水平Bcl-X-L的卵巢癌细胞系CaOV 3、SKOV 3 ipl和OVCAR 3中强制下调Bcl-X-L。我们还建立了过表达Bcl-X-L的OVCAR 3细胞的稳定转染子。我们最近表明Bcl-2调节卵巢癌细胞的细胞周期进程。因此，通过XTT测定法确定Bcl-X-L调节对细胞生长速率的影响。为了评估Bcl-X-L在药物诱导的细胞凋亡中的作用，在体外测定了三种细胞系中的每一种的顺铂和紫杉醇诱导的细胞凋亡。卵巢肿瘤细胞必须获得在非贴壁条件下生存的能力才能在腹水中生长。为了模拟锚定丧失并研究Bcl-X-L在此过程中的作用，将细胞培养在水凝胶包被的平板上，并通过核碎裂、caspase-3活化和核碘化丙啶染色来确定锚定丧失。我们发现，强制下调Bcl-X-L蛋白显着影响的CaOV 3的生长速率，而它对其他两个细胞系只有最小的影响。下调Bcl-X-L表达可增强CaOV 3、OVCAR 3和SKOV 3 ipl对顺铂和紫杉醇的敏感性。锚定的损失诱导的细胞凋亡的敏感性也增加，但在细胞系依赖性的方式。Bcl-XL过表达可抑制OVCAR 3细胞的生长，抑制药物诱导的凋亡和锚定丧失诱导的凋亡。总之，这些研究结果表明，Bcl-X-L的调节引起卵巢癌细胞生长和存活的变化是细胞系特异性的。因此，靶向Bcl-X-L的卵巢癌治疗策略可能会产生不同的反应。(c)2005年爱思唯尔公司All rights reserved.
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, SKOV3ipl 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 SKOV3ipl 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-XL 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. (c) 2005 Elsevier Inc. All rights reserved.