幽门螺杆菌感染对胃癌细胞m6A水平的影响及其机制

Effects of Helicobacter pylori infection on m6A level in gastric cancer cells and its mechanism

目的探讨幽门螺杆菌感染对胃癌中m6A水平的影响及机制。方法将胃癌细胞株MGC-803分别与幽门螺杆菌标准菌株NCTC 11637、NCTC 26695、PM SS1在MOI=100的条件下共培养8 h,用dot-blot和ELISA方法检测细胞总体m6A水平。用qRT-PCR和Western blotting检测METTL3和METTL14的表达。下载GEO数据库中的原始数据,分析m6A甲基化转移酶与临床病理参数之间的关系。运用GEPIA分析TCGA和GTEx数据库中m6A甲基化转移酶METTL14在胃癌和正常组织中的表达差异。结果幽门螺杆菌可导致胃癌细胞m6A水平上调;幽门螺杆菌感染诱导了胃癌细胞中METTL14表达;METTL14表达与胃癌组织LAUREN分型相关,且在肠型胃癌组织中表达更高;胃癌组织中METTL14的表达高于癌旁正常组织。结论幽门螺杆菌感染诱导胃癌细胞总体m6A水平上调,其机制与m6A甲基化转移酶METTL14表达上调有关。

Objective To investigate the effects of Helicobacter pylori(HP)infection on the level of m6A in gastric cancer and the mechanism.Methods After gastric cancer cell line MGC-803 was co-cultured with HP strains NCTC 11637,NCTC 26695 and PM SS1 at MOI=100 for 8 hours,the m6A level was detected with dot-blot and ELISA.The expressions of METTL3 and METTL14 were detected with qRT-PCR and Western blotting,respectively.Relevant data were downloaded from the GEO database to analyze the relationship between m6A methyltransferase and clinicopathological parameters.The difference in the expression of METTL14 in gastric cancer and normal tissues from TCGA and GTEx databases was analyzed with GEPIA.Results HP upregulated the m6A level in gastric cancer cells.HP infection promoted METTL14 expression.METTL14 expression correlated with LAUREN typing and was higher in intestinal type gastric cancer.METTL14 expression was higher in gastric cancer tissues than in normal adjacent tissues.Conclusion HP infection increases the m6A level in gastric cancer cells,which may be caused by the upregulation of METTL14.