目的 探讨隐丹参酮(CTS)对人胆管癌HCCC-9810细胞作用的敏感性并研究隐丹参酮对人胆管癌细胞周期及细胞凋亡率的影响,初步探讨隐丹参酮对人胆管癌细胞凋亡抑制基因Survivin表达的影响.方法 体外培养胆管癌细胞HCCC-9810,采用不同浓度隐丹参酮予以干预,通过细胞形态观察及MTT实验,初步研究隐丹参酮对人胆管癌细胞HCCC-9810的敏感性;应用流式细胞仪检测隐丹参酮诱导人胆管癌细胞周期的变化;Hochest染色法检测诱导细胞凋亡;RT-PCR检测人胆管癌HCCC-9810细胞Survivin基因mRNA表达变化.结果 隐丹参酮可明显抑制胆管癌HCCC-9810细胞株的增殖且该抑制效应随药物剂量的增加和作用时间的延长而增强;随着最适药物浓度作用时间的延长,胆管癌细胞明显被阻滞在G0/G1期,使S期细胞比例降低,细胞凋亡数目逐渐增加,Survivin基因mRNA表达逐渐降低.结论 隐丹参酮对人胆管癌HCCC-9810细胞增殖抑制作用呈剂量和时间依赖效应;隐丹参酮能诱导人胆管癌HCCC-9810细胞凋亡,其机制可能是通过影响肿瘤细胞的细胞周期、抑制Survivin基因表达有关.%Objective To investigate the sensitivity of cholangiocarcinoma HCCC-9810 cell lines to cryptotan-shinone; and to evaluate the effects of cryptotanshinone on cell cycle, apoptosis and survivin expression of cholangiocarcinoma cells. Methods HCCC -9810 cell lines were cultured in vitro and treated with cryptotanshinone of different concentrations. The sensitivity of cholangiocarcinoma cell(HCCC-9810)to cryptotanshinone was assessed with cell morphological observation and MTT method. Flow cytometry and RT-PCR were applied for evaluation of cell cycle and survivin mRNA expression, respectively, in HCCC -9810 cells. Results Cryptotanshinone significantly inhibited the proliferation of the HCCC -9810 cells following concentration and time dependent manners. As the optimal treatment spanned, HCCC -9810 cells was caught in G0/G1 phase, with reduced proportion of cells in S phase and increased apoptosis, and the survivin mRNA expression was down-regulated. Conclusion Cryptotanshinone can inhibit the growth of the HCCC-9810 following time and concentration dependent manners, Furthermore, cryptotanshinone induces apoptosis of HCCC-9810 cell, probably via down-regulation of survivin expression and affecting the cell cycle.
展开▼
