胃癌高发区幽门螺杆菌对胃上皮细胞系GES1蛋白质组的影响  

Effect of Helicobacter pylori isolated from the high incidence area of gastric cancer on the proteome of gastric epithelial cell line GES1

作　　者：魏思思[1] 邓晓晴 王苋 薛永娴 黄永胜 赵连梅[1] 单保恩[1] WEI Si-si;DENG Xiao-qing;WANG Xian;XUE Yong-xian;HUANG Yong-sheng;ZHAO Lian-mei;SHAN Bao-en(Research Center, the Fourth Hospital of Hebei Medical University, Shijiazhuang 050011;Shijiazhuang Center for Disease Control and Prevention, Shijiazhuang 050011;Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005, China)

机构地区：[1]河北医科大学第四医院科研中心,河北石家庄050011 [2]石家庄市疾病控制与预防中心,河北石家庄050011 [3]中国医学科学院基础医学研究所北京协和医学院基础学院,北京100005

出　　处：《基础医学与临床》2020年第12期1619-1625,共7页Basic and Clinical Medicine

基　　金：国家重点研发计划(2016YFC1302203);国家自然科学基金(81772550);河北省卫健委批准青年科技课题(20200113)。

摘　　要：目的探究胃癌高发区分离的幽门螺杆菌(H.pylori-400)对正常胃上皮细胞系GES1细胞蛋白质组的影响。方法C-13呼气实验阳性患者胃镜下取下胃黏膜,分离、培养并鉴定H.pylori-400。将H.pylori-400与GES1细胞按50∶1的比例共培养,提取细胞RNA,RT-qPCR法检测炎性因子IL-8、IL-1β和TNF-α的mRNA表达。收集共培养细胞蛋白,用高效液相色谱-质谱联用检测蛋白质组的改变,lable-free进行定量分析。选择差异表达蛋白进行GO和KEGG富集分析,并筛选出其中的关键基因(Hub)。结果在胃癌高发区(河北井陉地区)分离培养并鉴定出H.pylori-400,其能够促进胃上皮细胞GES1的IL-8,IL-1β和TNF-α炎性因子的mRNA表达。GES1感染H.pylori-400后,373个蛋白发生显著变化(|Fold change|≥2),其中143个蛋白降低,230个蛋白升高。这些差异表达蛋白经富集分析后表明,H.pylori-400可能通过调节细胞周期、T细胞免疫活性以及P53通路调节炎性因子分泌,进而导致肿瘤发生。结论胃癌高发区分离的H.pylori-400能够促进炎性因子表达,通过影响胃上皮细胞GES1蛋白质组而促进肿瘤发生。Objective To investigate the effect of Helicobacter pylori-400 that was isolated from high incidence area of gastric cancer on the proteome of gastric epithelial cell line GES1.Methods The gastric mucosa of patients with positive C-13 breath test was sampled with gastroscope,and H.pylori was isolated.GES1 was co-cultured with H.pylori-400 in the ratio of 1∶50,cellular RNA was extracted,and the mRNA expressions of IL-8,IL-1βand TNF-αwere detected by RT-qPCR.Cellular proteins were extracted and analyzed by HPLC-MS,lable-free was used for quantitative analysis.The differentially expressed proteins were selected for enrichment analysis of GO and KEGG,and the hub genes were screened out for further analysis.Results H.pylori-400 promoted mRNA expres-sion of IL-8,IL-1βand TNF-αof gastric epithelial cells.HPLC-MS showed that 373 proteins molecules changed(|Fold change|≥2)after GES1 co-cultured with H.pylori-400,among them,143 proteins molecules decreased and 230 protein molecules increased.GO and KEGG enrichment analysis showed that H.pylori-400 regulated the secretion of inflammatory factors by regulating cell cycle,T cell immune activity and p53 pathway,which may lead to tumor genesis.Conclusions H.pylori-400 isolated from the high incidence area of gastric cancer can promote the expression of inflammatory factors and promote the tumor genesis by affecting the proteome of GES1 cells.

关 键 词：幽门螺杆菌 胃上皮细胞 炎性因子 蛋白质组分析 

分 类 号：R37[医药卫生—病原生物学]