m6A甲基转移酶METTL3通过β‐arrestin 1/p65调控结肠癌细胞增殖  

m6A methyltransferase METTL3 regulates colon cancer cell proliferation throughβ⁃arrestin 1/p65

作　　者：江洁[1] 刘慧玲[1] 吴斌[1] JIANG Jie;LIU Hui-ing;WU Bin(Department of Gastroenterology,The Third Afffliated Hospital of Sun Yat-sen University,Guangzhou 510063,China)

机构地区：[1]中山大学附属第三医院消化内科,广东广州510063

出　　处：《解剖学研究》2024年第6期579-583,共5页Anatomy Research

摘　　要：目的探讨m6A甲基转移酶METTL3通过β-arrestin 1/p65在结肠癌细胞中的作用机制。方法使用结肠癌细胞系HCT116,应用荧光定量PCR、Western blot测定结肠癌细胞中METTL3、β-arrestin 1及增值蛋白PCNA、Cyclin D1及Cyclin E的表达水平;利用细胞转染及细胞克隆实验,观察METTL3、β-arres-tin1和p65在结肠癌细胞增殖中的作用。结果METTL3和β-arrestin 1在结肠癌细胞中表达升高(以actin作为内参基因,与正常对照组比较,P<0.05)。结肠癌细胞转染METTL3-shRNA后,转染组灰度(0.36±0.046)较对照组(1.27±0.14)明显减少(P=0.0004);与对照组克隆数(465±42)比较,转染组细胞增殖(85±37)明显减少(P<0.05);细胞增殖蛋白PCNA、Cyclin D1及Cyclin E的表达也明显减少(分别为P<0.01、0.05、0.01);结肠癌细胞中METTL3、β-arrestin 1和p-p65蛋白表达均增高(均为P<0.05)。随后HCT116细胞沉默MET-TL3的表达,发现细胞中β-arrestin 1、p-p65蛋白的表达明显下降(均为P<0.05)。结论METTL3影响结肠癌细胞的增殖可能通过β-arrestin 1/p65调控参与肿瘤的发生发展。Objective To investigate the mechanism of m6A methyltransferase METTL3 in colon cancer cells throughβ-arrestin 1/p65.Methods The expression of METTL3,β-arrestin 1,PCNA,Cyclin D1 and Cyclin E in colon cancer cell line HCT116 were determined by quantitative PCR and Western blot.The mechanism of MET-TL3,β-arrestin 1,p65 in the proliferation of colon cancer cells was studied by cell transfection and cell cloning ex-periments.Results The expression of METTL3 andβ-arrestin 1 was increased in colon cancer cells(With actin as the reference gene,compared with the normal control group,P<0.05).After transfection with shRNA,in cell clon-ing test,the number of clones was 465±42 in the control group and 85±37 in the transfection group(P<0.05).The ex-pression of cell proliferating protein PCNA,Cyclin D1 and Cyclin E was also significantly decreased(P<0.01,P<0.05 and P<0.01,respectively).In colon cancer cells,we found increased expressions of METTL3,β-arrestin1 and p-p65(All P<0.05).Subsequently,the expression of METTL3 was silenced in HCT116 cells,and the expression ofβ-arrestin1 and p-p65 in the cells was significantly decreased(All P<0.05).Conclusion METTL3 affects the prolifera-tion of colon cancer cells and may be involved in the development of cancer throughβ-arrestin1/p65 signaling pathway.

关 键 词：结肠癌 N6-甲基腺苷 甲基转移酶样蛋白3 β-抑制蛋白1 核因子κappa B 

分 类 号：R735.35[医药卫生—肿瘤]