摘要
本研究旨在比较、分析和验证东亚与西方幽门螺杆菌(Helicobacter pylori)菌株感染原代胃癌细胞后的差异表达基因(differentially expressed genes,DEGs),探讨东亚与西方菌株的致病差异。用幽门螺杆菌东亚菌株GZ7、西方菌株26695以感染复数(multiplicity of infection,MOI)50∶1感染原代胃癌细胞6 h,收取细胞进行转录组学测序,以|log_(2)FC|>1,FDR≤0.05为标准,筛选DEGs,利用OmicsBean-Cancer在线工具对DEGs进行生物信息学分析。选择4个重要基因进行TCGA数据库分析及Western blot验证。结果表明,GZ7感染胃癌细胞组与未感染胃癌细胞组比较有31个DEGs;26695感染胃癌细胞组与未感染胃癌细胞组比较有109个DEGs;26695感染组与GZ7感染组之间共有的DEGs为22个;GZ7感染组特有的DEGs为9个,26695感染组特有的DEGs为87个。DEGs主要与炎症(TNF信号通路、NF-κB信号通路)、细胞表面受体(Toll样受体信号通路、NOD样受体信号通路)等通路相关。TCGA数据库分析发现IL6和DKK1在胃癌组织中的表达显著高于正常组织的(P<0.001),且IL6和DKK1高表达与胃癌患者10年生存率呈负相关(P<0.01)。Western blot结果证实幽门螺杆菌感染上调胃癌细胞中JUN和FOSL1(两个转录因子),IL6,以及DKK1(东亚菌株特有的DEGs)蛋白的表达;下调细胞中的JUN基因可以抑制IL6和DKK1蛋白的表达,GZ7再感染能够恢复细胞中JUN、IL6和DKK1蛋白的表达水平。总之,东亚、西方幽门螺杆菌菌株感染胃癌细胞产生的DEGs有明显差异,这可能是东亚、西方国家胃癌发病率差异的原因之一。
This study aimed to compare,analyze,and verify the differentially expressed genes(DEGs)obtain from primary gastric cancer cells infected with East Asian and Western strains of Helicobacter pylori to understand the difference in pathogenic mechanisms between East Asian and Western strains of H.pylori.The primary gastric cancer cells were infected with the East Asian strain GZ7 and the Western strain 26695 of H.pylori for 6 h at multiplicity of infection(MOI)50∶1 and were collected for transcriptome sequencing.DEGs were screened with|log_(2)FC|>1 and FDR≤0.05.Bioinformatics analysis of the DEGs was performed using the online OmicsBean-Cancer tool.4 important genes of the DEGs were selected for further analysis by the TCGA database and validated by Western blot.There were 31 DEGs between the GZ7-infected and uninfected gastric cancer cells and 109 DEGs between the 26695-infected and uninfected gastric cancer cells groups,22 DEGs were shared between GZ7-and 26695-infected groups,9 DEGs were present only in GZ7 infected groups,and 87 DEGs present only in 26695-infected groups.These DEGs were involved in inflammation(TNF and NF-κB signaling pathway),cell surface receptors(Toll-like and NOD-like receptor signaling pathway)and so on.Analysis of the TCGA database revealed that IL6 and DKK1 in gastric cancer tissue were significantly higher than that in normal tissue(P<0.001),higher expression of IL6 and DKK1 was negatively associated with a 10-year survival rate of gastric cancer patients(P<0.01).Western blot result confirmed that H.pylori infection up-regulated the expression of protein JUN and FOSL1(two transcription factors),IL6 and DKK1(DEGs unique to East Asian strain)proteins in gastric cancer cells,and downregulation of JUN gene in cells suppressed the expression of IL6 and DKK1.When GZ7 was used to reinfect JUN-downregulated and control cells,protein expression of JUN,IL6 and DKK1 was restored.In conclusion,there is a clear difference in the DEGs generated by gastric cancer cells infected with the East Asian and Western strains of H.pylori which may be one of the reasons for the difference in the incidence of gastric cancer between East Asian and Western countries.
作者
廖永慧
龙妮娅
李雷蕾
彭国玲
陈源佳
谢渊
赵艳
周建奖
LIAO Yonghui;LONG Niya;LI Leilei;PENG Guoling;CHEN Yuanjia;XIE Yuan;ZHAO Yan;ZHOU Jianjiang(Key Laboratory of Ministry of Education for Endemic and Minority Diseases,Guizhou Medical University,Guiyang,550004;Key Laboratory of Molecular Biology,Guizhou Medical University,Guiyang,550004;Affiliated Hospital of Guizhou Medical University,Guiyang,550004)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2023年第2期121-132,共12页
Genomics and Applied Biology
基金
国家自然科学基金项目(32160166,31760328)
贵州省科技计划项目[黔科合支撑(2020)4Y026号,黔科合1Z010,黔科合基础(2020)1Y333]共同资助
关键词
幽门螺杆菌
胃癌
转录组测序
差异表达基因
Helicobacter pylori
Gastric cancer
Transcriptome sequencing
Differentially expressed genes
作者简介
通信作者:周建奖,zjj1985048@gmc.edu.cn
