索拉菲尼对肝癌细胞HepG2自噬的抑制作用

目的：研究化疗药物索拉菲尼（Sorafenib）对肝癌细胞HepG2自噬的作用及自噬与细胞增殖、细胞凋r_的关系。方法：常规细胞培养，以10μmol／L的索拉菲尼作用不同时间，采用MDC染色，在荧光显微镜下观察自噬泡的情况：用Western印迹检测自噬相关蛋白LC3的动态变化；用3-MA抑制肝癌细胞中自噬的表达，并用CCK8法检测索拉菲尼及其联合3-MA对细胞生存率的影响，用AnnexinV／PI流式细胞仪检测抑制自噬后凋亡的变化。结果：HepG2经索拉菲尼作用后，自噬泡明显减少，LC3蛋白尤其是LC3-Ⅱ随着作用时间延长逐渐减弱；索拉菲尼联合3-MA可进一步抑制HepG2细胞中自噬的表达，抑制自噬后细胞死亡明显增加，特别是细胞凋亡明显增加。结论：索拉菲尼抗肝癌作用可能与抑制肝癌细胞自噬有关。

Sorafenib inhibits human hepatoma cell line HepG2 autophagy

Objective To evaluate the effects of sorafenib on autophagy of human hepatoma cell line HepG2, and its relationship with cell proliferation and apoptosis. Methods Cell autophagy was observed under fluorescence microscopy by autophagosome staining with MDC. Autophagy-related protein LC3 expression was detected by western blot assay after the HepG2 cell line was treated with 10μM sorafenib for different period of time. The cellular viability was detected by the CCK8 method, and the apoptosis was detected by AnnexinV/PI flow cytometry after the cells were treated with sorafenib alone or combining with the autophagy inhibitor 3-MA. Results The autophagosomes were reduced significantly after sorafenib treatment, and LC3 protein （especially LC3-Ⅱ ） was concomitantly decreased. The autophagy was further inhibited by sorafenib in combination with 3-MA, acconlpanied by increased hepatocellular mortality and apoptosis. Conclusions Autophagy inhibition might play a role in the anti-tumor effects of sorafenib on human hepatic cancer