芹菜素通过上调PTEN蛋白表达诱导人肝癌细胞凋亡

Induction of apoptosis through up-regulating PTEN in human hepatocellular carcinoma HepG2 cell line by apigenin

目的观察芹菜素(apigenin,API)诱导人肝癌HepG2细胞凋亡的作用,研究其作用机制是否涉及PTEN蛋白表达的调控。方法体外培养HepG2细胞,PI染色流式细胞术分析凋亡率;琼脂糖凝胶电泳观察DNA梯形条带;WesternBlot分析细胞PTEN、p-Akt和p-Bad蛋白表达。结果API诱导HepG2细胞的凋亡,呈浓度依赖性。40μmol/L的API处理HepG2细胞48h,琼脂糖凝胶电泳呈现典型DNA梯形条带。40μmol/L的API处理HepG2细胞12h和24h,PTEN蛋白表达分别上调达132.6%和158.9%;同时磷酸化Akt蛋白水平下降至86.3%和75.6%,磷酸化Bad蛋白水平降低到73.4%和69.3%。结论API诱导HepG2细胞凋亡作用与上调PTEN蛋白表达、降低磷酸化Akt蛋白和磷酸化Bad蛋白水平相关。

[Objective] To investigate effects of Apigenin on apoptosis of human hepatocellular carcinoma HepG2 ceU line and its mechanism whether or not be involved in regulation of PTEN protein expression. [Methods] HepG2 cells were cultured in vitro. Flow cytometry using propidium iodide （PI） staining was used to determine cell apoptotic rate. DNA ladder bands were observed by DNA agarose gel electrophoresis. Western blot was used to analyze expression of PTEN, p-Akt and p-Bad proteins in HepG2 cells. [Results] FCM with PI staining demonstrated that Apigenin significantly induced the apoptosis of HepG2 cell line in a dose-dependant manner. DNA agarose gel electrophoresis showed that treatment of HepG2 cells with 40 μmol/L Apigenin for 48 h resulted in typical DNA ladder bands of DNA of HepG2 cells. Western blot analysis revealed that after 12 hours and 24 huors of treatment with 40 μmol/L apigenin, PTEN protein expression of HepG2 cells increased but p-Akt and p-Bad level decreased. [ Conclusion] Apigenin indueed apoptosis of human hepatocellular carcinoma HepG2 ceils by up-regulating PTEN and redueing phosphorylated Akt and Bad protein level.