Lin Fen, Yan Wei, Wen Ting, Wu Guo-yang
Department of General Surgery, Zhongshan Hospital, Xiamen University, Xiamen 361004, China.
Department of General Surgery, Zhongshan Hospital, Xiamen University, Xiamen 361004, China. Email:
Zhonghua Zhong Liu Za Zhi. 2013 Oct;35(10):742-6.
to investigate the effects of antidiabetic drug metformin on proliferation and apoptosis in human hepatocellular carcinoma cell line Huh-7 cells.
Huh-7 cells were treated with metformin at different concentrations. Cell viability was determined by MTT assay. Cell apoptosis and CD133(+) expression rate were detected by flow cytometery (FCM). Expressions of PTEN, Akt, p-Akt, Bcl-2, Bax proteins in the cells were measured by Western blot. The effect of metformin on the hepatosphere formation was observed in the serum-free suspension culture. Reverse transcription-polymerase chain reaction (RT-PCR) was used to validate the expression levels of stemness marker genes CD133, β-catenin, and ABCG2 mRNA in the hepatospheres.
The proliferation of Huh-7 cells was inhibited by metformin in a dose- and time-dependent manner. The early and late cell apoptosis rates induced by metformin at dose of 10 mmol/L for 48 hrs were (22.29 ± 0.8)% and (13.87 ± 1.2)%, respectively, and 25 mmol/L for 48 hrs (15.28 ± 2.1)% and (25.89 ± 2.3)%, respectively. Western blotting results revealed that the expression of CD133, phosphorylated Akt and the Bcl-2/Bax ratio were downregulated, and PTEN was upregulated in the Huh-7 cells after treated with 25 mmol/L metformin for 48 hrs. Metformin inhibited the formation of hepatospheres. Metformin also downregulated the expression of several cancer stem cells (CSCs)-related genes which are involved in the signaling pathways governing the self-renewal, proliferation and differentiation of CSCs in the hepatospheres.
Metformin inhibits the proliferation of human hepatocellular carcinoma Huh-7 cells and enhances their apoptosis in vitro. It may be related to the downregulation of PI3K/Akt signal pathway and selectively targeting CD133(+) cells.
研究抗糖尿病药物二甲双胍对人肝癌细胞系Huh-7细胞增殖和凋亡的影响。
用不同浓度的二甲双胍处理Huh-7细胞。采用MTT法测定细胞活力。通过流式细胞术（FCM）检测细胞凋亡和CD133(+)表达率。采用蛋白质免疫印迹法检测细胞中PTEN、Akt、p-Akt、Bcl-2、Bax蛋白的表达。在无血清悬浮培养中观察二甲双胍对肝球形成的影响。采用逆转录-聚合酶链反应（RT-PCR）验证肝球中干性标志物基因CD133、β-连环蛋白和ABCG2 mRNA的表达水平。
二甲双胍以剂量和时间依赖性方式抑制Huh-7细胞的增殖。10 mmol/L二甲双胍处理48小时诱导的早期和晚期细胞凋亡率分别为(22.29 ± 0.8)%和(13.87 ± 1.2)%，25 mmol/L二甲双胍处理48小时分别为(15.28 ± 2.1)%和(25.89 ± 2.3)%。蛋白质免疫印迹结果显示，25 mmol/L二甲双胍处理48小时后，Huh-7细胞中CD133、磷酸化Akt的表达及Bcl-2/Bax比值下调，PTEN上调。二甲双胍抑制肝球形成。二甲双胍还下调了几种癌症干细胞（CSCs）相关基因的表达，这些基因参与了肝球中CSCs自我更新、增殖和分化的信号通路。
二甲双胍在体外抑制人肝癌Huh-7细胞的增殖并增强其凋亡。这可能与PI3K/Akt信号通路的下调及选择性靶向CD133(+)细胞有关。
