川芎嗪抑制肝癌HepG2细胞增殖及对线粒体凋亡途径的影响

Inhibition of tetramethypyrazine on proliferation of HepG2 cells and its effects on the pathway of mitochondrial apoptosis

通过研究川芎嗪(TMP)诱导肝癌HepG2细胞凋亡,探讨TMP抑制肝癌细胞增殖的作用机制。采用CCK-8法检测TMP对肝癌HepG2细胞增殖的影响及量效关系,进一步应用高内涵细胞成像分析系统(HCS)检测TMP对HepG2诱导细胞凋亡作用以及对细胞色素C的释放和线粒体跨膜电位的影响,并结合Western blot检测TMP对肝癌HepG2细胞cleaved-caspase-3和cleaved-caspase-9表达的影响。结果显示,TMP对HepG2细胞增殖有抑制作用,并呈现剂量依赖性,可显著诱导HepG2细胞凋亡,增加细胞色素C从线粒体中释放到细胞质中,且能降低HepG2细胞内线粒体跨膜电位水平,以及显著提高肝癌HepG2细胞中cleaved-caspase-3及cleaved-caspase-9的表达水平。结果表明,TMP具有抑制肝癌HepG2细胞增殖的作用,其作用机制可能与通过线粒体途径触发细胞凋亡有关。

The purpose of the present study was to investigate the anti-proliferative and apoptotic effects of tetra- methypyrazine （TMP） on HepG2 and to elucidate the underlying mechanisms. CCK-8 was introduced to analyze the HepG2 cells proliferation. Cell apoptosis, mitochondrial membrane potential （△ψm） and cytochrome C were measured by high content screening （HCS）. Cleaved-caspases protein expression was detected by Western blot. CCK-8 assay indicated that TMP significantly inhibited HepG2 ceils proliferation in dose-dependent manner compared with the control group. Moreover, it was found that TMP could also induce HepG2 cell apoptosis, direct- ly increase the release of cytochrome C, decrease △ψm and increase cleaved-caspase-3 and cleaved-caspase-9 pro- tein expression. TMP may inhibit cell proliferation and induce cell apoptosis by stimulating the mitochondrial pathway apoptosis in HepG2 cells.