EGCG对人肝癌细胞的抑制作用及其可能的机制  

Epigallocatechin-3-gallate-induced growth inhibition and the underlying mechanisms in human hepatocellular carcinoma cells

作　　者：张勇[1] 沈筱芸[1] 冯雁[1] 谢裕安[1] 张力图[1] 利基林 罗小玲[1] 

机构地区：[1]广西医科大学附属肿瘤医院实验研究部,广西南宁530021

出　　处：《中国肿瘤生物治疗杂志》2014年第1期38-43,共6页Chinese Journal of Cancer Biotherapy

基　　金：广西医科大学青年科学基金项目(No.02602211011);广西自然科学青年基金项目(No.2013GXNSFBA019186);广西科学研究与技术开发计划课题(No.1140003A-32)~~

摘　　要：目的:观察表没食子儿茶素没食子酸酯[(-)-epigallocatechin-3-gallate,EGCG]对体外培养的人肝癌细胞株生物学特性的影响,研究其作用效果与血红素氧合酶-1(hemeoxygenase-1,HO-1)及相关信号分子的关系,探讨其作用机制。方法:利用MTT法检测EGCG对HepG2、Sk-hep1、SMMC7721等肝癌细胞增殖的影响,并用吖啶橙/溴化乙锭(AO/EB)双染法观察肝癌细胞的形态学变化,流式细胞术检测EGCG作用后Sk-hep1细胞周期的变化,Real-time PCR和Western blotting法检测EGCG作用后Sk-hep1细胞中HO-1、IL-10及TNF-α等信号分子表达的变化。结果:EGCG作用后,3株肝癌细胞贴壁细胞数量显著少于对照组,凋亡细胞数增多[HepG2:(16.33±3.51)vs(3.67±1.15)个,P<0.01),Sk-hep1:(18.33±2.31)vs(2.33±2.08)个,P<0.01),SMMC7721:(15.33±3.06)vs(3.33±2.08)个,P<0.01)]。实验组Sk-hep1细胞G2/M期比例明显高于对照组[(34.33±8.09)%vs(3.07±2.32)%,P<0.01]。设对照组基准值为1.00,实验组Sk-hep1细胞中HO-1、IL-10、及TNF-α的mRNA相对表达水平依次为(0.58±0.15)、(5.91±1.11)、(5.29±1.14),差别均有统计学意义(P<0.01);与对照组相比,实验组HO-1蛋白表达水平明显下调(0.16±0.04 vs 0.33±0.08,P<0.05),IL-10(0.42±0.06 vs 0.24±0.08,P=0.034,P<0.05)和TNF-α蛋白(0.95±0.17 vs 0.58±0.08,P<0.05)表达水平明显上调。结论:EGCG可抑制肝癌细胞增殖及诱导细胞凋亡,并将Sk-hep1细胞阻滞在G2/M期,其机制可能与HO-1、IL-10、TNF-α等炎症信号分子表达的变化有关。Objective ：To investigate the effect of Epigallocatechin-3-gallate （EGCG） on hepatocellular carcinoma cell growth and the molecular mechanisms underlying the effect in vitro . Methods： Three human hepatocellular carcinoma cell lines （i.e., HepG2, Sk-hep1 and SMMC7721） were used in this study. Cells were cultured in the presence of 0, 40, 80 or 120 μg/ml EGCG. At 24, 48 and 72 h after EGCG treatment, cell viability was assessed by MTT assay, apoptosis by AO/EB staining, cell cycle progression by flow cytometer, and mRNA and protein levels of HO11, IL-10 and TNF-α by Realtime PCR and Western blotting respectively. Results： EGCG treatment significantly induced cell attachment （ P 〈005）, increased the proportion of apoptotic cells （ P 〈0.01）, and induced G2/M arrest （ P 〈0.01） in all three cell lines tested as compared with the control. HO-1, IL-10 and TNF-α mRNA levels were 0.58±0.15, 5.91±1.11 and 529±1.14 in EGCG-treated Sk-hep1 cells, significantly different from the levels in the control cells （ P =0.008, P =0002, P =0.003）. EGCG resulted in a significant decrease in HO-1 protein content as compared with the control （0.16±0.04 vs 0.33±0.08, P 〈0.05）. In contrast, EGCG significantly increased levels of IL-10 protein （0.42± 0.06 vs 0.24±0.08, P 〈0.05） and TNF-α protein （0.95±0.17 vs 0.58±0.08, P 〈0.05）. Conclusions： EGCG may inhibit proliferation and block cell cycle progression and induce apoptosis in hepatocellular carcinoma cells. The mechanism（s） underlying these effects of EGCG may involve modulation of HO-1, IL-10 and TNF-α expression.

关 键 词：没食子儿茶素没食子酸酯 肝癌细胞 血红素氧合酶-1 

分 类 号：R735.7[医药卫生—肿瘤] R730.5[医药卫生—临床医学]