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

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.