吉林大学学报(医学版) ›› 2019, Vol. 45 ›› Issue (05): 1063-1068.doi: 10.13481/j.1671-587x.20190516

• 基础研究 • 上一篇    

幽门螺旋杆菌毒力因子CagA介导的ERK信号通路活化对胃癌细胞增殖和凋亡的影响

刘粉霞1, 陈丽1, 孙宁2, 吴亚薇1   

  1. 1. 河南省中医院病理科, 河南 郑州 450002;
    2. 河南中医药大学基础医学院, 河南 郑州 450002
  • 收稿日期:2018-10-30 发布日期:2019-10-08
  • 通讯作者: 刘粉霞,主治医师(Tel:0371-60905604,E-mail:fenxl88@163.com) E-mail:fenxl88@163.com
  • 作者简介:刘粉霞(1982-),女,河南省焦作市人,主治医师,医学硕士,主要从事胃癌致病机制方面的研究。
  • 基金资助:
    河南省卫计委科技攻关计划项目资助课题(162102310186,162102310331)

Effect of Helicobacter pylori virulence factor CagA-mediated activation of ERK signaling pathway on proliferation and apoptosis of gastric cancer cells

LIU Fenxia1, CHEN Li1, SUN Ning2, WU Yawei1   

  1. 1. Department of Pathology, Henan Provincial Hospital of Traditional Chinese Medicine, Zhengzhou 450002, China;
    2. School of Basic Medical Sciences, Henan University of Traditional Chinese Medicine, Zhengzhou 450002, China
  • Received:2018-10-30 Published:2019-10-08

摘要: 目的:探讨幽门螺旋杆菌毒力因子细胞毒素相关基因A蛋白(CagA)对细胞外调节蛋白激酶(ERK)信号通路的作用,阐明CagA的致癌机制。方法:构建pcDNA3.1/CagA真核表达载体,将胃癌AGS细胞分为空白对照组(空载体转染)、CagA转染组(GZ7/CagA转染)和CagA+ERKi组(ERK1/2抑制剂预处理+GZ7/CagA转染),采用Western blotting法测定各组细胞中CagA、磷酸化ERK(p-ERK)、总ERK(T-ERK)、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)和激活型caspase-3(cleaved caspase-3)蛋白表达水平,采用CCK-8法检测各组胃癌AGS细胞活性,采用流式细胞术检测胃癌AGS细胞凋亡率。结果:与空白对照组比较,CagA转染组胃癌细胞中CagA、p-ERK和Bcl-2蛋白表达水平明显升高(P<0.01),Bax和cleaved caspase-3蛋白表达水平明显降低(P<0.01);与CagA转染组比较,CagA+ERKi组胃癌细胞中p-ERK和Bcl-2蛋白表达水平明显降低(P<0.01),Bax和cleaved caspase-3蛋白表达水平明显升高(P<0.01);与CagA转染组比较,CagA+ERKi组各时间点胃癌细胞活性明显降低(P<0.01);与CagA转染组比较,CagA+ERKi组胃癌细胞凋亡率明显升高(P<0.05)。结论:幽门螺旋杆菌毒力因子CagA可抑制胃癌细胞凋亡,促进胃癌细胞增殖,其机制可能与CagA活化ERK信号通路有关。

关键词: 幽门螺旋杆菌, 细胞毒素相关基因A蛋白, 细胞外调节蛋白激酶, 胃癌细胞, 细胞增殖, 细胞凋亡

Abstract: Objective:To explore the effect of Helicobacter pylori virulence factor cytotoxin-associated gene A protein (CagA) on the extracellular regulated protein kinase (ERK) signaling pathway, and to elucidate the carcinogenesis mechanism of CagA. Methods:The pcDNA3.1/CagA eukaryotic expression vector was constructed, and the gastric cancer AGS cells were divided into blank control group (blank vector transfection), CagA transfection group (GZ7/CagA transfection), and CagA + ERKi group (ERK1/2 inhibitor pretreatment + GZ7/CagA transfection).The expression levels of CagA, phosphorylated ERK (p-ERK), total ERK (T-ERK), B-lymphocytoma-2 (Bcl-2), Bcl-2-related X protein (Bax) and cleaved caspase-3 proteins in the cells in various groups were determined by Western blotting method. The activities of AGS cells in various groups were determined by CCK-8 method, and the apoptotic rates of AGS cells were determined by flow cytometry. Results:Compared with blank control group, the expression levels of CagA, p-ERK, and Bcl-2 proteins in CagA transfection group were significantly increased(P<0.01), and the expression levels of Bax and cleaved caspase-3 proteins were significantly decreased (P<0.01). Compared with CagA transfection group, the expression levels of p-ERK and Bcl-2 proteins in CagA+ERKi group were significantly decreased (P<0.01), and the expression levels of Bax and cleaved caspase-3 proteins were significantly increased (P<0.01). Compared with CagA transfection group, the activities of gastric cancer cells in CagA + ERKi group at different time points were significantly decreased (P<0.01). Compared with CagA transfection group, the apoptotic rate of gastric cancer cells in CagA + ERKi group was significantly increased (P<0.05). Conclusion:Helicobacter pylori virulence factor CagA can inhibit the proliferation of gastric cancer cells and promote the apoptosis of gastric cancer cells, and its mechanism may be related to the activation of ERK signaling pathway by CagA.

Key words: Helicobacter pylori, cytotoxin-related gene A protein, extracellular regulated protein kinases, gastric cancer cells, cell proliferation, apoptosis

中图分类号: 

  • R735.2