KAT8通过增强METTL3表达促进结直肠癌细胞系增殖

KAT8 promotes the proliferation of colorectal cancer cell lines by enhancing METTL3 expression

目的探究赖氨酸乙酰转移酶8(KAT8)在结直肠癌细胞增殖过程中所扮演的角色及潜在作用机制。方法通过TCGA数据库的RNA-seq数据分析KAT8在结直肠癌患者的癌组织及癌旁组织的表达情况;集落形成实验和CCK8法检测KAT8敲低细胞的增殖;利用GEO数据库对KAT8敲低细胞与对照细胞的差异基因进行分析,并进一步进行功能通路富集分析;利用cBioPortal网站分析结直肠癌数据库中KAT8与调控N6-甲基腺苷(m6A)相关基因的相关性;Western blot检测KAT8和METTL3的蛋白表达。结果与人正常结直肠组织相比,KAT8在结直肠癌中高表达(P<0.05);敲低或选择性抑制KAT8抑制结直肠癌细胞系增殖(P<0.05);敲低KAT8降低结直肠癌细胞总RNA的m6A修饰水平(P<0.05);敲低KAT8抑制METTL3的表达(P<0.05);过表达METTL3能够逆转敲低KAT8抑制的细胞增殖(P<0.05)。结论KAT8通过增强METTL3介导的m6A修饰进而促进结直肠癌细胞系的增殖。

Objective To investigate the role and mechanism of lysine acetyltransferase 8(KAT8)on the prolifera⁃tion of colorectal cancer cells.Methods The expression of KAT8 in cancerous tissues and adjacent tissues of color⁃ectal cancer patients was analyzed by RNA⁃seq data of TCGA database.Cell proliferation was detected by colony⁃forming unit assays and CCK8.The GEO database was used to analyze the differential genes of KAT8 knockdown cells and control cells and perform functional pathway enrichment analysis.In the colorectal cancer database hosted on cBioPortal,that conducted an analysis examining the correlation between KAT8 and genes in the regulation of N6⁃methyladenosine(m6A)modification.Western blot technique was employed to assess the protein expression lev⁃els of KAT8 and METTL3.Results Compared to human normal colorectal tissue,KAT8 was highly expressed in colorectal cancer(P<0.05).Knockdown or selective inactivation of KAT8 inhibited colorectal cancer cells prolifera⁃tion(P<0.05).In colorectal cancer cell lines,knocking down KAT8 reduced m6A modification levels(P<0.05).Knocking down KAT8 inhibited METTL3 expression(P<0.05).Over⁃expression of METTL3 reversed cell prolifera⁃tion which was inhibited by knockdown KAT8(P<0.05).Conclusions KAT8 facilitates the proliferation of color⁃ectal cancer cell lines through regulation of METTL3⁃mediated m6A modifications.