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目的探讨环巴胺(cyclopamine,CYP)对人子宫内膜癌HEC-1A细胞存活及凋亡的影响。方法采用0,5,10,20,40μmol/L CYP处理HEC-1A细胞24 h,倒置显微镜下观察细胞形态改变,CCK-8法检测细胞增殖,AO/EB双染法观察死亡细胞;流式细胞仪AnnexinⅤ-FITC/PI法检测细胞发生的凋亡率,Q-PCR法检测Bax和Bcl-2基因的表达水平。结果 CYP处理导致HEC-1A细胞形态发生明显改变,且具有CYP浓度依赖性;CCK-8结果显示,CYP可显著抑制HEC-1A细胞的增殖(P<0.05);AO/EB染色结果表明,CYP可诱发HEC-1A细胞大量死亡;流式细胞术检测发现CYP可导致HEC-1A细胞凋亡,且随CYP浓度的升高HEC-1A细胞发生凋亡的比例升高(P<0.05);Q-PCR检测凋亡相关基因表达。结果显示,CYP处理可诱导HEC-1A细胞上调表达Bax基因,下调表达Bcl-2基因。结论 CYP可抑制人子宫内膜癌细胞HEC-1A的存活并诱导其发生凋亡。
Objective To investigate the effects of cyclopamine (CYP) on the survival and apoptosis of human endometrial carcinoma HEC-1A cells. Methods HEC-1A cells were treated with 0, 5, 10, 20 and 40μmol / L CYP for 24 hours. The morphological changes of the cells were observed under inverted microscope. Cell proliferation was detected by CCK-8 assay and apoptotic cells were observed by AO / Cell apoptosis rate was detected by Annexin V-FITC / PI method. The expression of Bax and Bcl-2 gene was detected by Q-PCR. Results The CYP treatment resulted in significant changes in the morphology of HEC-1A cells and a dose-dependent manner. The results of CCK-8 showed that CYP significantly inhibited the proliferation of HEC-1A cells (P <0.05). The results of AO / EB staining showed that CYP The apoptosis rate of HEC-1A cells increased with the increase of CYP concentration (P <0.05). Flow cytometry showed that CYP could induce the apoptosis of HEC-1A cells. Apoptosis-related gene expression was detected by PCR. The results showed that CYP treatment induced upregulation of Bax gene and down-regulation of Bcl-2 gene expression in HEC-1A cells. Conclusion CYP can inhibit the survival of human endometrial carcinoma cell HEC-1A and induce its apoptosis.