雷公藤甲素通过依赖于caspase的线粒体途径促进子宫内膜癌细胞凋亡

Triptolide induces caspase-dependent apoptosis in endometrial cancer via the mitochondrial pathway

目的:研究雷公藤甲素对子宫内膜癌HEC-1B细胞生长和凋亡的调节作用,探讨其促进子宫内膜癌细胞凋亡的作用途径。方法:①细胞增殖/毒性检测试剂(CCK-8)测定雷公藤甲素对HEC-1B细胞的生长抑制作用及加入caspase抑制剂(z-VAD-fmk)后对其作用的影响;②Western blot法检测用药后caspase-3、9和bcl-2蛋白表达情况及z-VAD-fmk对caspase-3、9蛋白表达的影响。结果:CCK-8比色法显示雷公藤甲素对子宫内膜癌HEC-1B细胞的生长有明显的抑制作用,且存在时间和浓度依赖性(P<0.01),在48、72 h作用效果明显。加入z-VAD-fmk后,雷公藤甲素对HEC-1B细胞生长抑制作用显著下降但仍高于对照组(P<0.01)。Western blot结果同样证明加入z-VAD-fmk后,caspase-3、9蛋白的表达有一定的减弱但仍高于对照组(P<0.05);同时随着药物浓度的增加,caspase-3、9蛋白表达逐渐升高,bcl-2蛋白表达逐渐降低(P<0.05)。结论:雷公藤甲素对子宫内膜癌HEC-1B细胞的生长有明显的抑制作用,通过依赖于caspase的线粒体凋亡途径促进子宫内膜癌HEC-1B细胞凋亡。

Objective：To investigate the pro-apoptosis effects of triptolide in human with endometrial cancer cell line （HEC-1B） on proliferation and apoptosis and its underlying mechanisms. Methods： ①The Cell Counting Kit-8 （CCK-8）was performed to measure the inhibitive effect of triptolide on the apoptosis on cell proliferation. ②The apoptosis and the change of caspase-3/9 in HEC-1B by pretreatment with the z-VAD-fmk （pan-capase inhibitor）were detected by CCK-8 and Western blot. Western blot was used to examine the expression of caspase-3/9,bcl-2. Results：The proliferation of HEC-1B was significantly decreased by triptolide in a dose-and time-dependent manner （P 〈 0.01）,and the effect in 48 h/72 h was obvious. The pan-caspase inhibitor z-VAD-fmk pretreatment reduced triptolide-induced apoptosis in HEC-1B, but the result was still higher than the control group （P 〈 0.01 ）. Triptolide increased the levels of caspase-3/9 expression while reduced bcl-2 expression in the dose-dependent manner （P 〈 0.05）. Conclusion： Triptolide functions as an effective apoptotic inducer in a caspase-dependent apoptosis via mitochondrial mechanism in human endometrial cancer （HEC-1B）.