β-榄香烯哌嗪衍生物DX2通过线粒体途径诱导人肝癌HepG2细胞凋亡

Piperazine derivate of β-elemene (DX2) induces human hepatoma HepG2 cell apoptosis through mitochondrial pathway

目的对β-榄香烯哌嗪衍生物DX2体外抑制人肝癌HepG2细胞增殖作用及其机制进行研究。方法采用MTT法测定细胞存活情况;采用倒置显微镜及吖啶橙染色观察细胞形态学变化;用流式细胞仪检测凋亡细胞比率及线粒体膜电位变化。用免疫印迹法分析DX2对细胞凋亡线粒体通路蛋白水平的影响。结果 DX2可明显抑制HepG2细胞增殖,诱导细胞出现凋亡小体,增加凋亡细胞比率。免疫印迹结果显示,35μmol/L DX2作用细胞后呈时间依赖性地降低HepG2细胞的线粒体膜电位,上调Bax/Bcl-2的比值,促进细胞色素c向胞浆释放,激活胱天蛋白酶(caspase)-9及其下游caspase-3,降解caspase-3底物ICAD。结论 DX2可诱导HepG2细胞凋亡,其机制为激活细胞凋亡线粒体途径。

Objective To study the antiproliferative effect of DX2,a piperazine derivate of β-elemene,on human hepatoma HepG2 cells. Methods Cell viability was measured by MTT method. Morphological changes were observed by phase contrast microscopy and acridine orange staining. Apoptotic cell ratio was measured by flow cytometric analysis. Protein level was detected by Western blot analysis. Results DX2 Induced apoptotic morphological changes in HepG2 cells and increased the ratio of apoptotic cells. Treatment of HepG2 cells with DX2 increased the Bax/Bcl-2 ratio,induced the activation of caspase-9 and caspase-3 and caused the degradation of ICAD which was the substrate of capase-3. Conclusion DX2 Induces HepG2 cell death via activation of intrinsic mitochondrial pathway.