TRIP13通过激活Notch1通路促进肝癌细胞对索拉非尼耐药的机制研究  

Mechanism of TRIP13 in promoting sorafenib resistance of hepatocellular carcinoma cells via activation of Notch1 signaling pathway

作　　者：陶冬英[1] 任典寰[1] 范任华[2] 杨菊林[1] 于纪棉[1] TAO Dongying;REN Dianhuan;FAN Renhua;YANG Julin;YU Jimian(Ningbo Vocational and Technical college,Ningbo 315806,Zhejiang,China)

机构地区：[1]宁波卫生职业技术学院,宁波315104 [2]长沙市中心医院

出　　处：《现代实用医学》2021年第5期571-574,614,共5页Modern Practical Medicine

基　　金：宁波卫生职业技术学院自然科学项目(2019Z02);宁波市科技局自然科学基金项目(2018A610384);浙江省科技厅公益类项目(LGD19H070001)。

摘　　要：目的探讨甲状腺激素受体相互作用蛋白13(TRIP13)在肝癌细胞中对索拉非尼耐药的作用和机制。方法采用浓度递增法建立HepG2索拉非尼耐药性细胞株HepG2-SR,real-time PCR和western blot检测HepG2亲本细胞和HepG2-SR细胞中TRIP13 mRNA和蛋白水平表达。然后将HepG2-SR细胞分成3组:对照组(加入10μM索拉非尼培养24 h)、si-NC组(转染50 nM si-NC 48h后加入10μM索拉非尼培养24 h)、si-TRIP13组(转染50 nM si-TRIP1348h后加入10μM索拉非尼培养24 h)。MTT检测索拉非尼对3组细胞的IC_(50),流式细胞术检测细胞凋亡,real-timePCR检测上皮间质转化基因E-cadherin和Vimentin表达,western blot检测Notch1通路活化相关蛋白Notch1和Hes-1表达水平。Starbase软件分析人肝癌组织中TRIP13和Notch1表达的相关性。结果HepG2-SR细胞中TRIP13 mRNA和蛋白水平均比HepG2亲本细胞高(均P<0.05)。在HepG2-SR细胞中,与si-NC组相比,si-TRIP13组中细胞的IC_(50)降低,凋亡细胞数目增多,E-cadherin表达增加,Vimentin、Notch1和Hes-1表达降低(均P<0.05)。在人肝癌组织中TRIP13和Notch1的表达呈正相关(r=0.33,P<0.01)。结论TRIP13在索拉非尼耐药的肝癌细胞中表达升高,其可促进肝癌细胞对索拉非尼的耐药这可能是通过激活Notch1通路,进而促进细胞的上皮间质转化发挥作用的。Objective To investigate the role and mechanism of TRIP13 in the sorafenib resistance of hepatocellular carcinoma cells.Methods HepG2 sorafenib resistant cell line(HepG2-SR)was established by incubating parental HepG2 cells with increasing concentrations of sorafenib.Real-time PCR and Western blot was used to detect the mRNA and protein levels of TRIP13 in HepG2 and HepG2-SR cells.Then,HepG2-SR cells were divided into three groups:control group(treatment with 10μM sorafenib for 24 h),si-NC group(transfection with 50 nM si-NC 48 h before 10μM sorafenib exposure)and si-TRIP13 group(transfection with 50 nM si-TRIP1348 h before 10μM sorafenib exposure).MTT assay was used to detect the IC50 of sorafenib.Cell apoptosis was detected by flow cytometry.Expression levels of epithelial mesenchymal transformation genes E-cadherin and Vimentin were determined by real-time PCR.Western blot was used to measure the levels of Notchl signaling activation-related proteins,including Notchl and Hes-1.Starbase software was used to analyze the correlation between TRIP13 and Notch1 in human hepatocellular carcinoma tissues.Results The mRNA and protein levels of TRIP13 were significantly higher in HepG2-SR cells than those in HepG2 parental cells(all P<0.05).In HepG2-SR cells,compared with the si-NC group,the IC50 decreased,and the number of apoptotic cells and Ecadherin expression increased,and the levels of Vimentin,Notchl,and Hes-1 decreased in si-TRIP13 group(all P<0.05).There was a positive correlation between the expression of TRIP13 and Notch1 in human hepatocellular carcinoma tissues(r=0.33,P<0.01).Conclusions TRIP13 is highly expressed in sorafenib resistant hepatocellular carcinoma cells.TRIP13 can promote sorafenib resistance of hepatocellular carcinoma cells,which may be through the activation of Notch1 pathway and subsequently promoting the epithelial mesenchymal transformation.

关 键 词：TRIP13 肝癌 索拉非尼耐药性 上皮间质转化 NOTCH1 

分 类 号：R735.7[医药卫生—肿瘤]