m^(6)A结合蛋白YTHDC2调节p38MAPK信号通路影响结直肠癌细胞凋亡  

The m^(6)A-binding protein YTHDC2 affects proliferation and apoptosis of colorectal cancer cells by regulating the p38MAPK signaling pathway

作　　者：张师垚 徐岩岩[1] 张琦[1] 李春强 赵智成[1] 刘刚[1] Zhang Shiyao;Xu Yanyan;Zhang Qi;Li Chunqiang;Zhao Zhicheng;Liu Gang(Department of General Surgery,Tianjin Medical University General Hospital,Tianjin 300052,China)

机构地区：[1]天津医科大学总医院普通外科,300052

出　　处：《中华结直肠疾病电子杂志》2023年第2期117-124,共8页Chinese Journal of Colorectal Diseases(Electronic Edition)

基　　金：天津市131创新型人才第一层次项目(303077900808);天津医科大学“十三五”学科建设项目(303071300302)。

摘　　要：目的探讨m^(6)A结合蛋白YTHDC2在结直肠癌发展过程中的分子机制及其所参与的信号通路。方法通过The Human Protein Atlas和GEPIA网站分析肿瘤基因组图谱(TCGA)数据库中YTHDC2与结直肠癌的关系,探究相关的信号转导途径和生物学过程。体外培养结直肠癌HCT116和Caco2细胞系,对YTHDC2进行过表达或敲低后,采用RT-PCR和Western blot检测p38MAPK、p-p38MAPK及其下游凋亡相关蛋白在丝裂原活化蛋白激酶(MAPK)信号通路中的表达,流式细胞术检测细胞凋亡率。结果The Human Protein Atlas和GEPIA分析结果表明,肿瘤组织中YTHDC2表达水平较癌旁组织存在降低趋势,YTHDC2的表达增加与结直肠癌患者总体生存率提高呈正相关。基于cBioPortal和Xena数据库中结直肠癌相关的基因表达量和临床数据以及KEGG和GO数据库中的注释基因集,基因集富集分析(GSEA)提示,YTHDC2具有调控MAPK信号通路的作用。流式细胞术检测结果显示,敲除YTHDC2时细胞凋亡比率显著减少,过表达时则显著增加,Western blot检测结果显示,p38MAPK表达未见显著变化,p-p38MAPK在YTHDC2过表达时显著升高,敲除时显著降低。基因表达谱交互分析(GEPIA)结果提示,凋亡蛋白的表达与YTHDC2的表达呈正相关,与RT-PCR和Western blot检测结果相符合。结论YTHDC2激活p38MAPK信号途径中外源性死亡受体和内源性线粒体凋亡通路调控结直肠癌细胞的凋亡。Objective To investigate the molecular mechanism and signal pathway of m^(6)A binding protein YTHDC2 in the development of colorectal cancer.Methods The relationship between YTHDC2 and colorectal cancer in the Cancer Genome Atlas(TCGA)database was analyzed through The Human Protein Atlas and GEPIA websites,and the related signal transduction pathways and biological processes were explored.Colorectal cancer cell lines HCT116 and Caco2 were cultured in vitro.After YTHDC2 was overexpressed or knocked down,the expression of p38MAPK,p-p38MAPK and their downstream apoptosis-related proteins in the mitogen-activated protein kinase(MAPK)signaling pathway was detected by RT-PCR and Western blot.The apoptosis rate was detected by flow cytometry.Results The Human Protein Atlas and GEPIA analysis results showed that the expression level of YTHDC2 in tumor tissues was lower than that in adjacent tissues,and the increase of YTHDC2 expression was positively correlated with the improvement of overall survival rate of patients with colorectal cancer.Based on the gene expression and clinical data related to colorectal cancer in cBioPortal and Xena databases and the annotated gene sets in KEGG and GO databases,gene set enrichment analysis(GSEA)suggested that YTHDC2 had a role in regulating the MAPK signaling pathway.The results of flow cytometry showed that the apoptosis rate was significantly reduced in YTHDC2 knockout group and increased in YTHDC2 overexpression group.Western blot showed that the expression of p38MAPK did not change significantly,while p-p38MAPK was significantly increased in YTHDC2 overexpression group and decreased in YTHDC2 knockout group.The results of gene expression profile interaction analysis(GEPIA)suggested that the expression of apoptotic proteins was positively correlated with the expression of YTHDC2,which was consistent with the results of RT-PCR and Western blot.Conclusions YTHDC2 activates exogenous death receptors and endogenous mitochondrial apoptosis pathways in the p38MAPK signaling pathway to

关 键 词：结直肠肿瘤 转录后修饰 细胞凋亡 YTHDC2蛋白 N-甲基腺苷 

分 类 号：R735.3[医药卫生—肿瘤]