GPCR48通过诱导肝癌细胞上皮间质转化促进肝癌侵袭转移的机制  

GPCR48 promotes invasion and metastasis by inducing epithelial-mesenchymal transition in hepatocellular carcinoma cells

作　　者：彭契六 韦尚谋 张磊 甘丽英 谢珍 覃举 张智[3] 韦进新 Peng Qiliu;Wei Shangmou;Zhang Lei;Gan Liying;Xie Zhen;Qin Ju;Zhang Zhi;Wei Jinxin(Department of Clinical Laboratory,Guangxi International Zhuang Medicine Hospital,Nanning 530201,China;Department of Clinical Laboratory,the Fifth Affliated Hospital of Guangxi Medical University&the First People's Hospital of Nanning,Nanning 530022,China;Department of Hepatobiliaty Surgery,the Fifth Affliated Hospital of Guangxi Medical University&the First People's Hospital of Nanning,Nanning 530022,China;department of Zhuang Medicine,Guangxi International Zhuang Medicine Hospital,Nanning 530201,China)

机构地区：[1]广西国际壮医医院检验科,南宁530201 [2]南宁市第一人民医院检验科,530022 [3]南宁市第一人民医院肝胆外科,530022 [4]广西国际壮医医院壮医综合病区,南宁530201

出　　处：《中华肝脏病杂志》2021年第9期849-854,共6页Chinese Journal of Hepatology

基　　金：广西自然科学基金面上项目(2017JJA10583);南宁市科学研究与技术开发计划项目(20163126);2019年广西中医药大学引进博士科研启动基金项目(2019BS042)。

摘　　要：目的观察G蛋白偶联受体48 (GPCR48)在不同转移潜能肝癌细胞株中的表达及其对肝癌细胞Huh7上皮间质转化(EMT)特性和侵袭转移的影响。方法采用蛋白质印迹(Western blot)检测不同转移潜能肝癌细胞GPCR48的蛋白表达水平。构建携带GPCR48基因的慢病毒载体,在肝癌细胞Huh7过表达GPCR48;采用Real-time PCR和Western blot检测GPCR48的过表达水平,采用Transwell侵袭实验和迁移实验检测对照组、Mock组和GPCR48过表达组肝癌细胞Huh7的侵袭和迁移能力;采用Real-time PCR和Western blot检测对照组、Mock组和GPCR48过表达组肝癌细胞Huh7 EMT相关标志物E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和γ连环蛋白(γ-catenin)的mRNA和蛋白的表达水平。数据组间差异比较采用方差分析。结果 GPCR48蛋白表达水平在转移性肝癌细胞株明显高于非转移性肝癌细胞株(P < 0.05)。携带GPCR48基因的慢病毒载体可以有效转染肝癌细胞Huh7,稳定表达GPCR48的mRNA和蛋白。GPCR48过表达组肝癌细胞Huh7与Mock组和对照组相比,侵袭和迁移能力明显增强(F≥5.54,P值均< 0.05),上皮表型标志物E-cadherin和γ-catenin的mRNA和蛋白表达水平下降(P值均<0.05),间质表型标志物N-cadherin和Vimentin的mRNA和蛋白表达水平上升(P值均< 0.05),表明过表达GPCR48的肝癌细胞Huh7发生了EMT改变。结论 GPCR48表达水平与肝癌细胞转移潜能呈正相关,过表达GPCR48可以下调上皮表型标志物的表达和上调间质表型标志物的表达,诱导肝癌细胞发生EMT改变,从而促进肝癌细胞侵袭和迁移。Objective To observe the G protein-coupled receptor 48(GPCR48)expression in hepatocellular carcinoma(HCC)cell lines with different metastatic potential and its characteristics effect on the invasion and metastasis of Huh7 hepatoma cells via epithelial-mesenchymal transition(EMT).Methods Western blot was used to detect the protein expression level of GPCR48 in HCC cells with different metastatic potential.The lentivirus vector expressing GPCR48 gene was constructed.GPCR48 was overexpressed in Huh7 hepatoma cells.The GPCR48 overexpression level was detected by real-time PCR and Western blot.Transwell invasion and migration assay was used to detect the Huh7 hepatoma cells invasion and migration ability in the Control,Mock and GPCR48 overexpression group.Real-time PCR and Western blot were used to detect Huh7 hepatoma cells mRNA and protein expression levels of the EMT related markers(E-cadherin,N-cadherin,vimentin,and y catenin)in the Control,Mock and GPCR48 overexpression groups,respectively.Analysis of variance was used to compare the dififerences between data sets.Results GPCR48 protein expression level in metastatic HCC cell lines was significantly higher than non-metastatic HCC cell lines(P<0.05).The lentivirus vector expressing the GPCR48 gene had effectively transfected the Huh7 hepatoma cells and stably expressed the GPCR48mRNA and protein.Compared with the Mock and the Control group,Huh7 hepatoma cells invasion and migration ability in the GPCR48 overexpression group was significantly enhanced(F≥5.54,P<0.05),and the mRNA and protein expression levels of epithelial phenotypic markers E-cadherin and y-catenin were decreased(P<0.05).The mRNA and protein expression levels of the mesenchymal phenotypic markers N-cadherin and Vimentin were increased(P<0.05),indicating that EMT changes occurred in Huh7 hepatoma cells had overexpressed GPCR48.Conclusion GPCR48 expression level is positively correlated with the metastatic potential of HCC cells.GPCR48 overexpression can down-regulate the expression of epithelial p

关 键 词：肝细胞癌 肿瘤转移 侵袭 上皮间质转化 G蛋白偶联受体48 

分 类 号：R735.7[医药卫生—肿瘤]