低浓度二甲双胍干预肝癌微环境诱导肿瘤细胞衰老的机制  

Studies on hepatocellular carcinoma cells senescence induced by low concentration of metformin via intervention of liver cancer microenvironment

作　　者：陈纪涛[1] 殷威达 刘兆宇[1] 李明[1] 方向明[1] 刘季芳[1] CHEN Ji-tao;YIN Wei-da;LIU Zhao-yu;LI Ming;FANG Xiang-ming;LIU Ji—fang(The Fifth Affiliated Hospital of Guangzhou Medical University,Guangzhou 510700,Guangdong,China)

机构地区：[1]广州医科大学附属第五医院肿瘤科

出　　处：《广东医学》2019年第24期3367-3371,共5页Guangdong Medical Journal

基　　金：国家自然科学基金面上项目(编号:81772553);广州市西医类一般引导项目(编号:20181A011090,20191A011089)

摘　　要：目的研究低浓度二甲双胍干预肝癌微环境诱导肿瘤细胞衰老抑制其增殖的机制。方法对5种肝癌细胞株分别采用0 mmol/L(对照组)、1 mmol/L(实验组)二甲双胍处理,筛选出高表达衰老相关β半乳糖苷酶(SA-β-Gal)的2种细胞株;通过MTS实验观察1 mmol/L二甲双胍对筛选出的肝癌细胞HepG2和Huh-7增殖的影响。采用免疫印迹检测低氧条件下药物干预HepG2和Huh-7细胞,钠氢交换蛋白1(NHE1)、质子转运ATP酶(V-ATPase)、HIF-1α、Glut1和CAXII蛋白的表达。结果肝癌HepG2和Huh-7细胞SA-β-Gal较对照组显著升高(P<0.05)。MTS结果显示二甲双胍抑制肝癌HepG2和Huh-7细胞的增殖。NHE1、V-ATPase、HIF-1α在对照组中表达高于实验组,而Glut1和CAXII蛋白两组相比无明显差异。结论低浓度二甲双胍可通过诱导肿瘤细胞衰老抑制其增殖,抗癌作用可能与改变肝癌细胞微环境pH值有关。Objective To investigate low doses of metformin intervention of liver cancer microenvironment by inducing hepatoma cells senescence and its underlying mechanism.Methods Five kinds of liver cancer cell lines were treated with 0 mmol/L(control group)and 1 mmol/L metformin(experimental group).Two cell lines with high expression of senescence-associatedβ-galactosidase(SA-β-Gal)were screened.The effect of metformin on the proliferation of selected hepatoma cells HepG2 and Huh-7 was observed by MTS assay.Immunoblotting was used to assess the expression of Na^+/H^+exchanger protein 1(NHE1),proton transport ATPase(V-ATPase),HIF-1α,Glut1 and CAXII proteins in HepG2 and Huh-7 cells.Results The expression of SA-β-Gal in HepG2 and Huh-7 were significantly increased in the liver cancer cells compared with the control group(P<0.05).MTS results showed that metformin inhibited the proliferation of HepG2 and Huh-7 cells.The expression of NHE1,V-ATPase and HIF-1αproteins was higher in the control group than in the experimental group,though there was no significant difference in Glut1 and CAXII proteins between the two groups.Conclusion Low concentration of metformin can inhibit the proliferation of tumor cells by inducing senescence,and may be correlated to the change of pH value of hepatocellular carcinoma microenvironment.

关 键 词：二甲双胍 肝细胞癌 钠氢交换蛋白1 质子转运ATP酶 衰老 

分 类 号：R735.7[医药卫生—肿瘤] R730.3[医药卫生—临床医学]