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

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.