PTEN低表达和c-Met高表达激活AKT/mTOR信号通路促进肝细胞癌形成  

The low expression of PTEN and along with the high expressions of c-Met activate AKT/mTOR signaling pathway and promote formation of hepatocellular carcinoma

作　　者：王瑞 戴琪 胡俊杰 WANG Rui;DAI Qi;HU Junjie(Laboratory of Pharmacology,School of Pharmacy,Hubei Univrsity of Traditional Chinese Medicine,Wuhan 430065,China;Department of Pharmacy,Huangiahu Hospital of Hubei University of Traditional Chinese Medicine,Wuhan 430065,China)

机构地区：[1]湖北中医药大学药学院药理实验室,湖北武汉430065 [2]湖北中医药大学校医院药剂科,湖北武汉430065

出　　处：《细胞与分子免疫学杂志》2021年第5期448-454,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：湖北省教育厅优秀中青年人才项目(Q20172001)。

摘　　要：目的探究第10号染色体缺失的磷酸酶和张力蛋白同源物(PTEN)低表达和细胞间质上皮转化因子(c-Met)高表达对肝细胞癌形成的影响及机制。方法实时定量PCR和免疫组织化学染色分别检测肝癌患者组织样本中PTEN和c-Met的mRNA和蛋白表达,并分析PTEN低表达和c-Met高表达患者的生存曲线。高压尾静脉注射法将提取制备的PTEN的短发夹RNA(shRNA)和c-Met基因质粒,单独或共同注射到FVB/N小鼠肝脏中,观察肿瘤的发生发展情况并绘制各组小鼠生存曲线,Western blot法检测小鼠肝组织蛋白激酶B/哺乳动物雷帕霉素靶蛋白(AKT/mTOR)信号通路的激活。雷帕霉素不敏感性mTOR伴侣分子基因敲除(Rictorflox/flox)小鼠分别转染注射shPTEN/c-Met/PT3(对照组)、shPTEN/c-Met/Cre(Rictor敲除组),观察肿瘤的发生和发展情况,Western blot法检测各组小鼠肝组织AKT、mTOR蛋白的磷酸化水平。结果在60例肝癌样本中,44例样本PTEN表达量低于癌旁组织,42例的c-Met的表达高于癌旁组织,PTEN低表达和c-Met高表达同时发生在34例患者中。shPTEN质粒和c-Met基因质粒共同注射组FVB/N小鼠肝癌发生和死亡率显著高于shPTEN质粒和c-Met基因质粒单独注射组,AKT/mTOR信号通路激活情况亦显著高于shPTEN质粒和c-Met基因质粒单独注射组。Rictorflox/flox小鼠转染shPTEN协同c-Met基因质粒肝癌发生率及AKT、mTOR蛋白的磷酸化均显著低于转染shPTEN协同c-Met基因质粒的野生型小鼠。结论肝癌中PTEN低表达和c-Met高表达可促进AKT/mTOR信号通路的激活而促进肝癌的发生。Objective To explore the effects of the low expression of phosphatase and tensin homolog deleted on chromosome ten(PTEN)along with the high expression of c-Mesenchymal-epithelial transition factor(c-Met)on the formation of hepatocellular carcinoma,and study the related mechanism.Methods Real-time quantitative PCR and immunohistochemistry were used to detect the expression of PTEN and c-Met in tissue samples of liver cancer patients,and then the survival curve of patients with low PTEN expression and high c-Met expression was analyzed.The high-pressure tail vein injection method was used to inject the extracted PTEN shRNA and c-Met gene plasmid into the liver of FVB/N mice individually or together.The occurrence and development of tumors were monitored and survival curves were drawn in each group.Western blotting was performed to detect the activation of AKT/mTOR signaling pathway in mouse liver tissue.Rapamycin-insensitive companion of mTOR knockout mice(Rictorflox/flox mice)were injected with shPTEN/c-Met/PT3(control group),shPTEN/c-Met/Cre(Rictor knockout group),and the occurrence and development of the tumors were observed,and the phosphorylation levels of AKT and mTOR proteins in the liver of each group were evaluated by Western blotting.Results The real-time quantitative PCR and immunohistochemical detection showed that PTEN expression in 44 tumor sampleswas lower than that in the adjacent tissues among 60 samples,and c-Met expression in 42 tumor samples was higher than that in the adjacent tissues.The low expression of PTEN and the high expression of c-Met occurred in 34 patients.The incidence of liver cancer and the mortality in the mice injected with shPTEN plasmid and c-Met gene plasmid were significantly higher than those in shPTEN plasmid and c-Met gene plasmid alone transfection group,and the activation of AKT/mTOR signal pathway was also significantly higher than that of shPTEN plasmid and c-Met plasmid transfected separately.The incidence of liver cancer and the phosphorylation level of AKT and mTOR

关 键 词：第10号染色体缺失的磷酸酶和张力蛋白同源物(PTEN) 细胞间质上皮转化因子(c-Met) 肝癌 蛋白激酶B(AKT) 哺乳动物雷帕霉素靶蛋白(mTOR) 

分 类 号：R735.7[医药卫生—肿瘤] R392-33[医药卫生—临床医学]