DNA甲基转移酶N6AMT1基因敲降和过表达及其功能初探  

Knock down and over-expression of the DNA methyltransferase gene N6AMT1 and preliminary study for its functions

作　　者：杨钰[1] 孙振[1] 薛松磊[1] 胡序明[1] 崔恒宓[1,2] YANG Yu;SUN Zhen;XUE Songlei;HU Xuming;CUI Hengmi(Epigenetics and Epigenetic Institute of Genomics/College of Animal Science and Technoloy,Yangzhou University,Yangzhou 225009,China;Joint International Research Laboratory of Agriculture&Agri-Product Safety,Ministry of Education,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]扬州大学动物科学与技术学院/表观遗传学及表观基因组学研究所,江苏扬州225009 [2]扬州大学教育部农业与农产品安全国际合作联合实验室,江苏扬州225009

出　　处：《扬州大学学报（农业与生命科学版）》2019年第5期26-32,共7页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家自然科学基金资助项目(81773013);国家重点研发计划项目(2016YFC1303604)

摘　　要：6mA甲基化是DNA甲基化修饰的一种重要方式,受甲基转移酶N6AMT1基因的调控。为研究N6AMT1基因的功能,利用CRISPR/Cas9基因编辑技术构建N6AMT1基因敲降的HEK293细胞系,同时在Hela细胞系中过表达N6AMT1基因,检测N6AMT1基因表达变化对细胞增殖和迁移能力的影响。结果表明:正常肾上皮组织细胞系HEK293中N6AMT1基因表达水平显著高于癌细胞;正常肝细胞系LO2中N6AMT1基因表达水平显著高于肝癌细胞系HepG2和SMMC-7721;N6AMT1基因敲降显著增强HEK293细胞的增殖及迁移能力;而Hela细胞过表达N6AMT1基因后,细胞的增殖和迁移均显著受到抑制。这一研究提示N6AMT1基因介导的6mA甲基化可能对细胞增殖和迁移有重要影响。6 mA methylation is an important DNA methylation modification which is regulated by methyltransferase gene N6AMT1. In order to study the functions of N6AMT1 gene, a N6AMT1 knockdown HEK293 cell line was constructed by CRISPR/Cas9 editing technique. At the same time, N6AMT1 was over-expressed in Hela cell line. The effect of N6AMT1 expression on cell proliferation and migration was detected. The results showed that the expression level of the N6AMT1 gene in normal renal epithelial cell line HEK293 and normal hepatocyte line LO2 was significantly higher than that in renal cancer cells, hepatocellular carcinoma cell line HepG2 and SMMC-7721;the knockdown of the N6AMT1 gene significantly enhanced the proliferation and migration of HEK293 cells;and the proliferation and migration of Hela cells were significantly inhibited after over-expression of N6AMT1. These results suggest that DNA 6 mA methylation mediated by N6AMT1 may play an important role in cell proliferation and migration.

关 键 词：DNA 6mA甲基化 N6AMT1基因 CRISPR/Cas9 肿瘤 

分 类 号：Q78[生物学—分子生物学]