幽门螺杆菌感染对胃上皮细胞m^(6)A甲基化水平及相关酶表达的影响  

Effects of Helicobacter pylori infection on m^(6)A methylation level and related enzymes expression in gastric epithelial cells

作　　者：桂书琴 何小凤 吴丽霞 赵艳[1,2] 王琴容[1,2] 周建奖[1,2] 谢渊[1,2] Gui Shuqin;He Xiaofeng;Wu Lixia;Zhao Yan;Wang Qinrong;Zhou Jianjiang;Xie Yuan(Key Laboratory of Endemic and Ethnic Minority Diseases,Ministry of Education,Guizhou Medical University,Guiyang 550004;Key Laboratory of Molecular Biology,Guizhou Medical University,Guiyang 550004)

机构地区：[1]贵州医科大学地方病与少数民族性疾病教育部重点实验室,贵阳550004 [2]贵州医科大学分子生物学重点实验室,贵阳550004

出　　处：《安徽医科大学学报》2023年第10期1639-1645,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:31960028、32160166);贵州省科学技术基金(编号:黔科合基础[2020]1Z010、黔科合基础[2020]1Y333)。

摘　　要：目的研究幽门螺杆菌(Hp)感染胃上皮细胞中N6-甲基腺苷(m^(6)A)修饰水平及相关酶FTO、METTL3和YTHDF2的表达变化,并使用在线网站分析其在胃癌中的表达和临床意义。方法Hp GZ7菌株感染正常胃上皮细胞GES-1和胃癌细胞AGS,感染复数为30∶1,24 h后收集细胞总RNA和总蛋白,斑点杂交实验检测细胞总RNA的m^(6)A修饰水平,q-PCR和Western blot检测FTO、METTL3和YTHDF2的mRNA和蛋白表达。GEPIA网站分析FTO、METTL3和YTHDF2在胃癌组织和正常组织中的表达差异及其在Stage分期中表达,Kaplan-Meier Plotter分析FTO、METTL3和YTHDF2表达与预后的关系。结果与未感染组比较,Hp感染GES-1和AGS细胞中总RNA m^(6)A修饰水平均明显降低,FTO的mRNA和蛋白表达水平增加,METTL3和YTHDF2的mRNA和蛋白水平降低,差异有统计学意义(P<0.05)。胃癌组织中FTO表达水平高于正常组织,差异有统计学意义(P<0.05),YTHDF2表达水平高于正常组织,METTL3表达水平低于正常组织,差异无统计学意义。FTO和METTL3表达较高的胃癌患者有较低的生存率,而YTHDF2高表达患者有较高的生存率。FTO表达与患者的Stage分期显著相关(P<0.05),而METTL3与YTHDF2表达与Stage分期无显著相关性。结论Hp感染可引起细胞m^(6)A水平及相关酶FTO、METTL3和YTHDF2表达改变,提示m^(6)A甲基化可能在Hp诱导胃癌发生发展中发挥作用。Objective To investigate the expression of N6-methyladenosine modification(m^(6)A)and related enzymes FTO,METTL3 and YTHDF2 in gastric epithelial cells infected with Helicobacter pylori(Hp),and to analyze its expression and clinical significance in gastric cancer by using online websites.Methods Gastric epithelial cells GES-1 and AGS were infected with Hp GZ7 strain,and the multiplicity of infection was 30∶1.Total RNA and total protein were collected after infecting 24 h,and the m^(6)A modification level of total RNA was detected by RNA dot hybridization.The mRNA and protein expressions of FTO,METTL3 and YTHDF2 were detected by q-PCR and Western blot.The expression difference of FTO,METTL3 and YTHDF2 in gastric cancer tissues and normal tissues and their expression in Stage classification were analyzed by GEPIA,and the relationship between the expression of FTO,METTL3 and YTHDF2 and prognosis was analyzed by Kaplan-Meier Plotter.Results[WTBZ]Compared with the uninfected group,the m^(6)A modification level of total RNA in both Hp-infected GES-1 and AGS cells significantly decreased,the mRNA and protein expression levels of FTO increased,and the mRNA and protein levels of METTL3 and YTHDF2 decreased,with statistical significance(P<0.05).The expression level of FTO in gastric cancer tissues was higher than that in normal tissues,with statistical significance(P<0.05),YTHDF2 was higher than that in normal tissues,with no statistical significance,and the expression level of METTL3 was lower than that in normal tissues,with no statistical significance.Patients with high expression of FTO and METTL3 had a lower survival rate,while patients with high expression of YTHDF2 had a higher survival rate.The expression of FTO was significantly correlated with Stage classification(P<0.05),while the expression of METTL3 and YTHDF2 was not significantly correlated with Stage classification.Conclusion Hp infection can change the level of m^(6)A and the expression of related enzymes FTO,YTHDF2 and METTL3,suggesting that m^(6)A meth

关 键 词：胃癌 幽门螺杆菌 m^(6)A FTO METTL3 YTHDF2 

分 类 号：R735.2[医药卫生—肿瘤]