丙氨瑞林与DDP对卵巢癌耐药细胞CoC1/cDDP凋亡的影响

目的 探讨促性腺激素释放激素类似物(GnRHa)丙氨瑞林(Alarelin)与顺铂(DDP)对人卵巢癌耐药细胞CoC1/cDDP凋亡的影响.方法 CoC1/cDDP细胞分别加入不同浓度的丙氨瑞林及DDP+丙氨瑞林培养,设空白对照组(不加药),流式细胞仪检测亚G1期细胞及凋亡细胞和线粒体膜电位(ΔΨm),半定量RT-PCR法检测CoC1/cDDP细胞生存素(survivin)-⊿Ex3 mRNA及天冬氨酸特异性半胱氨酸蛋白酶(caspase-3 mRNA的表达.采用四甲基偶氮唑蓝(MTT)法测定各组细胞生长抑制率.结果 ①随DDP浓度增加,亚G1期细胞及AnnexinV+/PI-细胞比例上升,对细胞的抑制作用增强(P＜0.05).丙氨瑞林作用于卵巢癌耐药细胞CoCl/cDDP后,随浓度增加降低ΔΨm,亚G1期细胞及AnnexinV+/PI-细胞比例增加,凋亡细胞增多,细胞增殖抑制率上升(P＜0.01).联合应用丙氨瑞林及10 μg/ml DDP后,线粒体膜电位(ΔΨm)降低,Rh123-细胞数增加,G1期细胞及AnnexinV+/PI-细胞增多,细胞抑制率明显上升,强于单用DDP或丙氨瑞林；②丙氨瑞林单独或联合DDP作用于CoCl/cDDP细胞,caspase-3mRNA的表达随丙氨瑞林浓度增加而上调,但survinin-⊿ Ex3 mRNA表达无明显变化(P＞0.05).结论 丙氨瑞林能促进凋亡,逆转CoCl/cDDP细胞对DDP耐药,其机制可能与降低ΔΨm,上调caspase-3mRNA表达有关.

The effects of Alarelin combined with DDP on apoptosis in the drug resistant human ovarian cancer cell Line CoC1/cDDP

Objective To investigate the effect and the mechanisms of Alarelin combined with cisplatin（DDP）on the apoptosis in drug-resistant human ovarian cancer cell line CoC1/cDDP.Methods CoC1/cDDP cell were treated with different concentrations of alarelin,DDP and alarelin＋DDP.The survivin-Ex3mRNA,caspase-3 mRNA were measured by RT-PCR.The G1cells and apoptosis cells and the cell mitochondrial membrane potential（ΔΨm）were measured by FCM.The growing inhibiting rates were examined by MTT.Results ① The apoptosis cells and the growing inhibiting rates increased with the concentration of DDP increased（P0.05）.Alarelin increased the percentages of sub-G1and AnnexinV＋/PI-cells in a dose-dependent manner（P0.01）.Compared with separate use alarelin or DDP,after treating with alarelin＋10μg/ml DDP,the apoptosis cells increased more obviously.The percentages of sub-G1,AnnexinV＋/PI-cells and the percentages of Rh123cells were increased（P 0.01）.②Alarelin significantly increased the expression of caspase-3mRNA in a dose-dependent manner（P 0.01）,while treating with alarelin or alarelin combined with DDP,the expression of survivin-Ex3mRNA had no significant increased（P0.05）.Conclusions Alarelin has strong reversal effects on DDP resistance in CoC1/cDDP,the effects may be related to decreasing theΔΨm and increasing expression of caspase-3mRNA.