FH535抑制人肝癌细胞HepG2增殖及cyclin D表达

Inhibitory effect of FH535 on proliferation and cyclin D expression in HepG2 cell line

目的探讨β-catenin抑制剂FH535对人肝癌细胞系HepG2增殖的影响及可能机制。方法 HepG2细胞分为对照组和FH535用药组,使用改良3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐比色法(MTS)检测FH535对HepG2细胞增殖的影响,以Western blot法检测HepG2细胞β-catenin以及cyclin D蛋白表达水平,用实时荧光定量聚合酶链式反应(PCR)检测HepG2细胞cyclin D mRNA水平。结果 FH535能够显著抑制HepG2细胞的增殖,呈时间和剂量依赖性。与对照组相比,FH535给药组HepG2细胞内β-catenin蛋白表达无差异。FH535给药组细胞wnt/β-catenin信号通路的靶基因cyclin D的mRNA和蛋白的表达显著下降,与对照组相比差异有统计学意义(P<0.0001)。结论 FH535通过抑制cyclin D mRNA及cyclin D蛋白表达而抑制HepG2细胞增殖。

Objective To explore the effect of β-catenin inhibitor FH535 on proliferation of HepG2 and its mechanism.Methods HepG2 cells were treated with FH535.Cell proliferation was determined by modified 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide colorimetric assay（MTS）.cyclin D mRNA expression were determined by real time polymerase chain reaction（RT-PCR）,β-catenin and cyclin D protein expression were determined by western blot.Results Compared with the control group,the proliferation of HepG2 cells was significantly decreased after treatment with FH535,in a dose-and time-dependent manner.cyclin D mRNA and cyclin D protein level were both down-regulated in FH535 group compared with the control group（P0.0001）,while β-catenin protein expression was not affected.Conclusions FH535 inhibited the proliferation of HepG2 cells.This might through down-regulation of cyclin D mRNA and cyclin D protein.