二甲双胍激活AMPK促进顺铂对肝癌细胞凋亡的初步研究

Prelimary studies on metformin activation of AMPK to make cisplatin induce hepatocellular carcinoma cells apoptosis

目的探讨二甲双胍与顺铂合用对肝癌细胞的作用,为进一步的体内研究奠定基础。方法 MTT法检测不同浓度、不同时间二甲双胍、顺铂单用及两者合用于SMMC7721、HepG2肝癌细胞所致细胞增殖及凋亡;Hoechst33258检测二甲双胍、顺铂单用及合用对HepG2细胞核形态变化的影响。Western blot检测细胞内AMPK/p-AMPK的表达。结果二甲双胍、顺铂单用及合用于SMMC7721、HepG2,随着作用时间及浓度增加抑制率增加,呈时间-浓度依赖性;Hoechst33258结果显示合用组细胞凋亡最严重;Western blot结果显示二甲双胍、顺铂单用及合用均激活AMPK,联合用药组p-AMPK上调最明显。结论二甲双胍联合低浓度顺铂可激活AMPK,增加其对SMMC7721、HepG2癌细胞的毒性。

[ Objective ] To investigate the effect of mefformin combined with cisplatin （DDP） on the hepatocellu- lar carcinoma cells. [ Methods ] The effect of metformin combined DDP on the growth of human hepatic carcinoma cell line HepG2, SMMC7721 in vitro was evaluated by MTY assay. The Morphological changes of nucleus of HepG2 were evaluated by Hoechst33258 staining. The expression of AMPK/p-AMPK was evaluated by Western blot. [Re- sults ] With the time and dosage increasing of metformin, cisplatin alone or combination, the inhibition for HepG2 and SMMC7721 was more effective. Cell apoptosis was the most apparent in combination group by Hoechst33258. AMPK was activated by metformin, cisplatin alone or combination, the upregulation of p-AMPK in the combination group was the most obvious. [ Conclusion ] Metfoimin combined cisplatin can activate AMPK to increase the cytotoxicity for tumor cell.