幽门螺杆菌CagA通过B-Raf和JNK2调节原癌蛋白CIP2A表达及其细胞生物学作用研究  

Helicobacter pylori CagA upregulates the expression of CIP2A by B-Raf and JNK2 and its cellular biological function

作　　者：张亚云 徐智渊 赵大鹏[3] 王玉[2] 焦凤萍[4] 杨庆玺[5] 庄东明[4] 李晓霞[2] 孙铮 赵英会[2] ZHANG Ya-yun;XU Zhi-yuan;ZHAO Da-peng;WANG Yu;JIAO Feng-ping;YANG Qing- xi;ZHUANG Dong-ming;LI Xiao-xia;SUN Zheng;ZHAO Ying-hui(Anesthesiology, theFirst Hospital Affiliated with Zhengzhou University, Zhengzhou 450052, China;Department of Pathogen Biology, College of Basic Medical Sciences, Taishan Medical University;Clinical Laboratory, Taishan Medical University Hospital;School of Public Health, Taishan Medical University;Internal Medicine, Taishan Convalescent Hospital of Shandong Province;The Second School of Clinical Medicine, Taishan Medical University)

机构地区：[1]郑州大学第一附属医院麻醉科,河南郑州450052 [2]泰山医学院病原生物学教研室 [3]泰山医学院附属医院检验科 [4]泰山医学院公共卫生学院 [5]山东省泰山疗养院内科 [6]泰山医学院第二临床医学院

出　　处：《中国病原生物学杂志》2017年第12期1148-1151,1156,共5页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.81602455);山东省自然科学基金项目(No.ZR2013HM033;ZR2013HL057;ZR2013HL060);泰山医学院重大科研专项(No.2014GCC10);泰安市科研课题(No.2015NS2098;2016NS1074;2016NS1089)

摘　　要：目的了解幽门螺杆菌CagA上调CIP2A表达的分子机制。方法培养AGS细胞和GES-1细胞,一部分细胞分别转染CagA阳性质粒WT-cagA和阴性对照质粒pcDNA3.1;另一部分细胞用B-Raf和JNK2信号抑制剂作用,然后分别转染质粒WT-cagA和质粒pcDNA3.1,Western blot检测CIP2A表达,细胞克隆形成试验检测细胞增殖能力,细胞迁移试验检测细胞迁移能力结果 AGS细胞和GES-1细胞转染CagA阳性质粒WT-cagA后CIP2A表达量增多;细胞经B-Raf和JNK2的信号抑制剂作用后转染CagA阳性质粒WT-cagA,其CIP2A表达量比仅转染质粒WT-cagA的细胞表达量少,细胞的克隆数减少,细胞迁移能力降低。结论幽门螺杆菌CagA进入细胞通过B-Raf和JNK2及其参与的信号途径调节原癌蛋白CIP2A的表达,进而影响细胞的克隆形成能力和迁移能力。Objective To ascertain the molecular mechanism by which Helicobacter pylori CagA upregulates CIP2A. Methods After AGS and GES-1 cells were separately cultured to exponential phase,AGS or GES-1 cells were separately transfected with either the recombinant plasmid WT-cagA or the control vector plasmid pcDNA3.1.The cells were then treated with a signal inhibitor of B-Raf or JNK2 and transfected with the plasmid recombinant WT-cagA.The expression of CIP2A was detected with Western blotting,and the cell clonogenic potential was determined using a cell cloning assay.Cell migration was detected with a cell migration assay. Results AGS and GES-1 cells transfected with the plasmid WT-cagA had a higher level of CIP2A expression than that in the control.After treatment with a signal inhibitor of B-Raf or JNK2 and subsequent transfection with the plasmid WT-cagA,the level of CIP2A expression in the cells was decreased.The cell clonogenic potential and cell migration decreased below the cell clonogenic potential and cell migration noted in cells transfected with the plasmid WT-cagA alone. Conclusion H.pylori CagA upregulates the expression of the oncogene CIP2A through B-Raf and JNK2,affecting the cell clonogenic potential and cell migration.

关 键 词：幽门螺杆菌 胃癌 CAGA CIP2A B-RAF JNK2 

分 类 号：R378.91[医药卫生—病原生物学]