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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 study the regulatory effect of triptolide on the growth and apoptosis of endometrial carcinoma HEC-1B cells and to explore its role in promoting the apoptosis of endometrial cancer cells. (1) Cell proliferation / toxicity assay (CCK-8) was used to determine the effect of triptolide on the growth of HEC-1B cells and the effect of triptolide plus its caspase inhibitor (z-VAD-fmk) The expression of caspase-3, 9 and bcl-2 protein and the effect of z-VAD-fmk on the expression of caspase-3 and 9 proteins were detected. Results: CCK-8 colorimetry showed that triptolide significantly inhibited the growth of endometrial carcinoma HEC-1B cells in a time and concentration dependent manner (P <0.01) obvious. After adding z-VAD-fmk, the inhibitory effect of triptolide on HEC-1B cells was significantly decreased but still higher than that of the control group (P <0.01). The result of Western blot also showed that the expression of caspase-3, 9 was decreased but still higher than that of the control group (P <0.05) after the addition of z-VAD-fmk. At the same time, The expression of bcl-2 protein gradually decreased (P <0.05). Conclusion: Triptolide significantly inhibits the growth of endometrial carcinoma HEC-1B cells and promotes the apoptosis of HEC-1B cells through a caspase-dependent mitochondrial apoptotic pathway.