Wang Haixia, Zhang Zhifeng, Wei Xiuping, Dai Ruizhen
J Ovarian Res. 2015 Feb 20;8:3. doi: 10.1186/s13048-015-0130-x.
Bcl-2 plays a major role in the pathobiology and drug resistance of ovarian cancer, and inhibition of bcl-2 was useful for OC therapy. It has previously reported that TW-37, a small-molecule inhibitor of Bcl-2 family proteins, inhibited cell growth and induced apoptosis in many cancer cells. In the present study,we investigate the effect of TW-37 or / and in combination with cisplain on several ovarian cancer (OC) cell lines with high bcl-2 expression.
The bcl-2 mRNA and protein expression, and the cisplain (DDP) sensitivity of OC cell lines SKOV3, OVCAR3, OV-90 and 3AO and SKOV3DDP were determined by Quantitative real-time RT-PCR,Western blot, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and fluorescence-activated cell sorting (MTT) assays. The effects of TW-37 alone or combined with cisplain on growth and apoptosis in bcl-2 overexpressed OVCAR3, OV-90 and SKOV3DDP cells was detected by MTT,clonogenic assay, ELISA and terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay.
The cell lines SKOV3 and 3AO were sensitive, whereas OVCAR3, OV-90 and SKOV3DDP were resistant to cisplain. Significant positive correlation was observed between basal bcl-2 mRNA and protein and cisplain sensitivity. Cisplain treatment did not activate bcl-2 in vitro. Treatment with TW-37 inhibited bcl-2 expression in bcl-2 overexpressed OVCAR3, OV-90 and SKOV3DDP cells , and inhibited growth and induced apoptosis ,and increased cisplain killing of the bcl-2 overexpressed cells in a does and time-dependant manner in vitro.
Bcl-2 level positively correlated with sensitivity to cisplain. Treatment with TW-37 was effective alone and in combination with cisplain in bcl-2 overexpressed OC cell lines in vitro. Thus, TW-37 may be a useful therapeutic agent for OCs.
Bcl-2在卵巢癌的病理生物学和耐药性中起主要作用，抑制Bcl-2对卵巢癌治疗有益。此前有报道称，TW-37作为一种Bcl-2家族蛋白的小分子抑制剂，可抑制多种癌细胞的生长并诱导其凋亡。在本研究中，我们研究了TW-37单独或与顺铂联合使用对几种高表达bcl-2的卵巢癌细胞系的影响。
通过定量实时逆转录聚合酶链反应、蛋白质免疫印迹法以及噻唑蓝和荧光激活细胞分选法检测卵巢癌细胞系SKOV3、OVCAR3、OV-90、3AO和SKOV3DDP的bcl-2信使核糖核酸和蛋白表达以及对顺铂的敏感性。通过噻唑蓝、克隆形成试验、酶联免疫吸附测定和末端脱氧核苷酸转移酶介导的缺口末端标记法检测TW-37单独或与顺铂联合使用对bcl-2过表达的OVCAR3、OV-90和SKOV3DDP细胞生长和凋亡的影响。
细胞系SKOV3和3AO对顺铂敏感，而OVCAR3、OV-90和SKOV3DDP对顺铂耐药。基础bcl-2信使核糖核酸和蛋白与顺铂敏感性之间存在显著正相关。顺铂处理在体外未激活bcl-2。用TW-37处理可抑制bcl-2过表达的OVCAR3、OV-90和SKOV3DDP细胞中bcl-2的表达，抑制细胞生长并诱导凋亡，且以剂量和时间依赖的方式增加顺铂对bcl-2过表达细胞的杀伤作用。
Bcl-2水平与对顺铂的敏感性呈正相关。在体外，TW-37单独使用以及与顺铂联合使用对bcl-2过表达的卵巢癌细胞系均有效。因此，TW-37可能是一种治疗卵巢癌的有效药物。
