大黄素甲醚调节miR-370诱导肝癌细胞凋亡的实验研究

Regulatory effect of physcion on induced apoptosis in hepatocellular carcinoma by miR-370

目的:观察大黄素甲醚调节miR-370诱导肝细胞癌(HCC)细胞的凋亡情况,并探讨其作用机制。方法:将大黄素甲醚作用于SMMC7721和HepG2两组HCC细胞,使用TaqMan探针用实时定量PCR法检测miR-370的表达;用Western blot检测Sp1和DNMT1相应的蛋白表达水平。结果:大黄素甲醚抑制肝细胞癌细胞增长和诱导凋亡。肝细胞癌细胞中通过上调miR-370造成大黄素甲醚诱导型细胞凋亡。大黄素甲醚处理的细胞中通过miR-370抑制剂抑制miR-370水平能明显降低凋亡细胞的百分比(P<0.01)。大黄素甲醚通过AMPK/Sp1/DNMT1信号调节miR-370的水平(P<0.01)。AMPK/Sp1/DNMT1信号参与大黄素甲醚诱导的HCC细胞凋亡。结论:大黄素甲醚通过上调miR-370诱导HCC细胞凋亡,通过调节AMPK/Sp1/DNMT1信号通路对miR-370发挥调节作用。

Objective： To observe and investigate the mechanism of induced apoptosis of hepatic cell carcinoma by miR-370. Methods： HCC cell lines （SMMC7721 and HepG2） were treated with physcion. MiR-370 expression was quantified by real time PCR with a TaqMan Probe and mRNA expression of Spl and DNMT1 was quantified by Western blot, Results： Physcion suppressed cell growth of HCC cells and increased the apoptotic proportion in both cell lines. Physcion induced apoptosis in HCC cells through upregulating miR-370. Our findings showed that the physcion modulated the level of miR-370 through AMPK/Sp1/DNMT1 signaling （P〈0.01）. Conclusions： Physcion exerts anti-tumor effect against HCC, and may be a potential agent for the adjunct chemotherapy.