CerS2对膀胱癌细胞增殖、迁移及AKT/mTOR信号通路的影响  

Effects of CerS2 on bladder cancer cell proliferation,migration and AKT/mTOR signaling pathway

作　　者：吕建阳[1] 李振国 陈林[1] 刘殿刚[2] LYU Jianyang;LI Zhenguo;CHEN Lin;LIU Diangang(Department of Urology,Beijing Sixth Hospital,Beijing 100007,China;General Surgery,Xuanwu Hospital,Capital Medical University,Beijing 100053,China)

机构地区：[1]北京市第六医院泌尿外科,北京100007 [2]首都医科大学宣武医院普通外科,北京100053

出　　处：《现代肿瘤医学》2021年第22期3885-3889,共5页Journal of Modern Oncology

基　　金：北京市卫生系统高层次卫生技术人才培养计划项目(编号:2015-3-066)。

摘　　要：目的:探究CerS2基因在膀胱癌细胞中的作用及其可能的机制。方法:构建pEGFP-C1-CerS2重组表达载体,并转染膀胱癌细胞系T24,使用蛋白质免疫印迹实验检测GFP-CerS2的表达;运用CCK-8法检测细胞增殖活性;运用Annexin V荧光标记实验检测细胞凋亡水平;分别使用细胞划痕实验及Transwell实验检测细胞迁移能力和侵袭能力;使用蛋白质免疫印迹实验检测细胞中AKT、p-AKT、mTOR及p-mTOR的表达水平。结果:重组GFP-CerS2融合蛋白在T24细胞中可以正常表达,过表达GFP-CerS2显著抑制了膀胱癌细胞的增殖活性,且显著增加了细胞凋亡水平。过表达GFP-CerS2抑制了膀胱癌细胞的迁移和侵袭能力。此外,过表达GFP-CerS2也显著抑制了膀胱癌细胞中AKT和mTOR的磷酸化修饰,但不影响总AKT及mTOR的表达水平。结论:过表达CerS2可以抑制AKT/mTOR信号通路,影响膀胱癌细胞的增殖和凋亡,并抑制细胞的迁移和侵袭能力。Objective:To investigate the role and the possible mechanisms of CerS2 in bladder cancer cells.Methods:The reconstitution expression vectors of pEGFP-C1-CerS2 was constructed,and then transfected into T24 bladder cancer cells.The expression levels of GFP and GFP-CerS2 were examined by immunoblotting assay.The cells proliferation activities were measured by CCK-8 assays.The apoptosis rates were measured by Annexin V fluorescent labeling.The cells migration activities and invasion activities were measured respectively by wound healing assay and Transwell assay.The expression levels of AKT,p-AKT,mTOR and p-mTOR were measured by immunoblotting assay.Results:The reconstituted GFP-CerS2 fusion protein was well expressed in T24 cells.Overexpression of GFP-CerS2 significantly inhibited the proliferation activities in bladder cancer cells,and increased the apoptosis rate.Overexpression of GFP-CerS2 also inhibited the migration and invasion capabilities in bladder cancer cells.In addition,overexpression of GFP-CerS2 remarkably decreased the phosphorylation of AKT and mTOR,but did not affect the total expression of AKT and mTOR.Conclusion:Overexpression of CerS2 suppresses AKT/mTOR signaling pathway,promotes apoptosis,and inhibits the migration and invasion capabilities in bladder cancer cells.

关 键 词：CerS2 膀胱癌 增殖 凋亡 迁移 AKT/MTOR 

分 类 号：R737.14[医药卫生—肿瘤]