摘要
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).
出处
《南京医科大学学报(自然科学版)》
CAS
CSCD
北大核心
2013年第2期168-171,共4页
Journal of Nanjing Medical University(Natural Sciences)
基金
江苏省中医药局科技基金资助(HZ07029)
江苏省肿瘤医院重点专科建设项目(ZS201202)