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[目的]观察过氧化物酶体增殖物激活受体γ(PPARγ)的配体罗格列酮(RSG)对肝癌细胞凋亡的影响。[方法]体外培养人肝癌HepG2细胞,将HepG2细胞分为对照组和RSG不同浓度(25、50、100、200μmol·L-1)加药组。应用流式细胞仪检测RSG不同浓度对肝癌细胞HepG2凋亡率的影响;应用Hoechst33258荧光染色观察细胞凋亡形态学变化;实时荧光定量PCR方法观察Bcl-2、Bax的mRNA表达变化;免疫细胞化学法检测Bcl-2、Bax、PPARγ的蛋白表达。[结果]流式细胞仪和荧光染色法显示RSG诱导肝癌HepG2细胞凋亡,凋亡率与浓度呈正相关;RSG干预后,Bcl-2的mRNA及蛋白在肝癌细胞中表达下调,Bax的mRNA及蛋白在肝癌细胞中表达均上调。[结论]RSG依赖激活PPARγ能在体外诱导肿瘤细胞凋亡,其机制可能是通过下调抗凋亡基因Bcl-2表达以及上调促凋亡基因Bax而实现的,提示PPARγ可能是肝癌治疗的一个新分子靶点。
[Objective] To observe the effect of rosiglitazone (RSG), a ligand of peroxisome proliferator activated receptor γ (PPARγ), on the apoptosis of hepatocellular carcinoma cells. [Methods] HepG2 cells were cultured in vitro. HepG2 cells were divided into control group and RSG groups (25, 50, 100, 200 μmol·L-1). Flow cytometry was used to detect the effect of different concentrations of RSG on the apoptotic rate of HepG2 cells. Hoechst33258 fluorescence staining was used to observe the morphological changes of apoptotic cells. Real-time fluorescence quantitative PCR was used to observe the mRNA expression of Bcl-2 and Bax. Immunocytochemistry Method to detect the protein expression of Bcl-2, Bax and PPARγ. [Results] Flow cytometry and fluorescence staining showed that RSG induced apoptosis in HepG2 cells. The apoptosis rate was positively correlated with the concentration of RSG. After RSG intervention, the mRNA and protein of Bcl-2 were down-regulated in hepatocellular carcinoma cells, The protein was up-regulated in hepatoma cells. [Conclusion] RSG-dependent activation of PPARγ can induce tumor cell apoptosis in vitro. The mechanism may be through down-regulating the expression of anti-apoptotic gene Bcl-2 and up-regulating the pro-apoptotic gene Bax, suggesting that PPARγ may be a new treatment for hepatocellular carcinoma Molecular target.