姜黄素对人肝母细胞瘤细胞株HepG2内β-连环蛋白信号通路的影响  

Effects of curcumin on β-catenin signaling pathways in human hepatoblastoma cell line HepG2

作　　者：李勇[1] 肖雅玲[1] 陈朝晖[1] 黎明[1] 李青玲[1] 邹欣[1] 周海燕[2] 

机构地区：[1]湖南省儿童医院普外科,长沙410007 [2]中南大学基础医学院病理学系

出　　处：《中华小儿外科杂志》2014年第7期535-539,共5页Chinese Journal of Pediatric Surgery

摘　　要：目的观察姜黄素对体外培养HepG2中p连环蛋白（β-catenin）信号通路的影响，探讨姜黄素抑制肝母细胞瘤的分子机制。方法实时定量聚合酶链反应（RT-PCR）及免疫印迹法（WesternBlot）检测对照组及不同浓度姜黄素干预组（10、20、30、40、50μmol／L）HepG2中β-cateninmRNA及蛋白表达水平；TOPflash／FOPflash双荧光素酶报告系统检测β-catenin的转录活性；RThPCR检测各组pcatenin靶基因细胞周期素D1（cyclinD1）、血管内皮细胞生长因子（VEGF）及基质金属蛋白酶9（MMP9）mRNA表达水平。结果HepG2高表达β-cateninmRNA及蛋白，各浓度姜黄素干预均能抑制HepG2细胞pcateninmRNA表达（P〈0．05或P〈0．01），10-50μmol／L姜黄素干预分别降低了β-cateninmRNA表达水平20％、40％、48％、58％和60％；20～50μmol／L姜黄素干预均能抑制cyclinD1、VEGF及MMP9mRNA的表达（P〈0．05或P〈0．01）；20-50μmol／L姜黄素干预均能抑制HepG2细胞pcatenin蛋白表达（P〈O．05或P〈0．01），分别降低30％、43％、60％和75％；20-50μmol／L姜黄素干预分别抑制HepG2中β-catenin的转录活性27％、36％、55％和60％（P〈0．05或P〈0．01）。结论姜黄素可能通过抑制β-catenin的表达及转录活性而影响pcatenin信号通路相关下游靶基因的表达，这可能是其发挥抗肿瘤效应的分子机制之一。Objective To explore the effects of curcumin （cur） on β-catenin signaling pathways in HepG2 and elucidate its anti-tumor molecular mechanism. Methods The expressions of mRNA and protein of β-eatenin in control HepG2 and cureumin treatment groups of different concentrations （10, 20, 30, 40, 50/~mol/L） were evaluated by reverse transcription-polymerase chain reaction （RT-PCR） and Western blot. And the transcriptional activity of β-eatenin was assayed by TOP flash/FOP flash dual-luciferase reporter systerm The expressions of β-catenin targeted gene cyclin D1, VEGF and MMP9 were evaluated by RT-PCR. Results β-catenin was highly expressed in HepG2. Curcumin （10-50 vmol/L） could inhibit the mRNA expression of β-catenin by approximately 20%, 40%, 48 %, 58% and 60% respectively. Meanwhile curcumin （20-50 μmol/L） significantly inhibited the mRNA expression of β-eatenin targeted gene MMP9, eyclin D1 and VEGF （P〈0. 05 or P〈0. 01）. The expression of β-catenin at the protein level decreased by approximately 30%, 43%, 60G and 75% after treatment. And the transcriptional activity of β-catenin in HepG2 was also inhibited by different concentrations of cureumin （20-50 μmol/L） and decreased by approximately 27%, 36%, 55% and 60% respectively （P〈 0. 05 or P〈0. 01 ）. Conclusions Curcumin can inhibit the expression and transcriptional activity of β-eatenin and affect the expression of β-eatenin targeted gene through the β- catenin signaling pathways. And it may be a possible mechanism of its anti-neoplastic effect.

关 键 词：肝肿瘤 姜黄素 Β-连环蛋白 细胞周期素D1 

分 类 号：R735.7[医药卫生—肿瘤]