大黄素对大鼠肝癌细胞凋亡诱导因子及核酸内切酶G mRNA表达的影响

Effects of Emodin on mRNA Expression of Apoptosis-inducing Factor and Endonuclease G of Rat CBRH-7919 Cells

目的研究大黄素(emodin,EM)对大鼠肝癌细胞CBRH-7919细胞凋亡及凋亡诱导因子(apoptosis inducing factor,AIF)和核酸内切酶G(endonuclease G,EndoG)mRNA表达的影响,探讨大黄素的抗癌机制。方法实验分空白对照组、Caspase抑制剂Z-VAD-FMK干预组、大黄素组、大黄素+Z-VAD-FMK组共4组,采用四氮唑盐(MTT)还原法筛选大黄素的干预条件,根据结果选择44.8μmol/L的大黄素干预48h作为干预条件;以原位末端标记法(Tunel)检测细胞凋亡;实时荧光定量PCR(real-time quantitative PCR,QT-PCR)法检测AIF、EndoGmRNA。结果大黄素能抑制肝癌CBRH-7919细胞株的增殖,诱导其凋亡,与空白组相比差异具有显著性(P<0.01);QT-PCR检测显示,大黄素及大黄素+Z-VAD-FMK组细胞AIF、EndoG mRNA表达上调,与空白组比较,差异具有显著性(P<0.01)。结论大黄素可上调CBRH-7919细胞AIF、EndoG表达,通过启动非Caspase依赖通路而发挥促凋亡的作用。

Objective To observe the effects of emodin on mRNA expression of apoptosis - inducing factor （AIF） and endonuclease G （EndoG） in rat CBRH - 7919 cells, and to investigate its mechanism in inducing apoptosis. Methods The CBRH - 7919 cells were divided into 4 groups： blank control group, caspase inhibitor Z - VAD - FMK group, emodin group, emodin combined with Z - VAD - FMK group. In - situ end labeling method （TUNEL） were used to detect the apoptosis of CBRH - 7919 cells. The mRNA expression of AIF and EndoG were examined by real - time quantitative PCR. Results Emodin obviously increased the apoptosis rate of rat CBRH - 7919 cells in vitro （ P 〈 0.01 compared with the blank group）. The mRNA expression of AIF and EndoG were significantly increased in emodin group and emodin combined with Z - VAD - FMK group （ P 〈 0.01 compared with the blank group）. Conclusion Emodin could induce the apoptosis of CBRH - 7919 cells in vitro by upregulating the expression of AIF and EndoG, which can induce apoptosis through the caspase - independent pathway.