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

基础医学与临床 ›› 2020, Vol. 40 ›› Issue (12): 1619-1625.

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

魏思思¹, 邓晓晴¹, 王苋², 薛永娴¹, 黄永胜³, 赵连梅¹, 单保恩^1*

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

收稿日期:2020-06-08 修回日期:2020-09-30 出版日期:2020-12-05 发布日期:2020-11-30
通讯作者: ^* shanbaoen@163.com
基金资助:
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)

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

WEI Si-si¹, DENG Xiao-qing¹, WANG Xian², XUE Yong-xian¹, HUANG Yong-sheng³, ZHAO Lian-mei¹, SHAN Bao-en^1*

1. Research Center, the Fourth Hospital of Hebei Medical University, Shijiazhuang 050011;
2. Shijiazhuang Center for Disease Control and Prevention, Shijiazhuang 050011;
3. Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005, China

Received:2020-06-08 Revised:2020-09-30 Online:2020-12-05 Published:2020-11-30
Contact: ^* shanbaoen@163.com

摘要/Abstract

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。

关键词: 幽门螺杆菌, 胃上皮细胞, 炎性因子, 蛋白质组分析

Abstract: 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-1β and TNF-α were 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-1β and TNF-α of 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.

Key words: Helicobacter pylori, gastric epithelial cell, inflammatory factors, proteome

中图分类号:

魏思思, 邓晓晴, 王苋, 薛永娴, 黄永胜, 赵连梅, 单保恩. 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[J]. 基础医学与临床, 2020, 40(12): 1619-1625.

WEI Si-si, DENG Xiao-qing, WANG Xian, XUE Yong-xian, HUANG Yong-sheng, ZHAO Lian-mei, SHAN Bao-en. Effect of Helicobacter pylori isolated from the high incidence area of gastric cancer on the proteome of gastric epithelial cell line GES1[J]. Basic & Clinical Medicine, 2020, 40(12): 1619-1625.

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