JARID2对牛卵丘细胞H3K9me3、H3K27me3的甲基化修饰作用

Effect of JARID2 on H3K9me3 and H3K27me3 methylation in bovine cumulus cells

本试验利用RNAi方法抑制牛卵丘细胞中JARID2基因mRNA的表达水平,通过免疫荧光方法检测H3K4me3、H3K9me3、H3K27me3、H3K36me3的甲基化程度。结果表明,在牛卵丘细胞中存在JARID2的表达,并且存在H3K9me3、H3K27me3的甲基化修饰,但没有检测到H3K4me3、H3K36me3的甲基化修饰。通过siRNA对牛卵丘细胞中JARID2 mRNA的表达进行成功抑制后,转染组JARID2的表达量明显下降,但H3K4me3、H3K27me3表达量无明显变化,而H3K9me3、H3K27me3表达量明显下降,即JARID2在mRNA水平上能够影响H3K9me3、H3K27me3的甲基化修饰程度,而对H3K4me3、H3K36me3的甲基化修饰无明显作用。

Jmj family members JHDM1,JHDM2 Aare capable of specifically removing H3K9 and H3K36methylation modification,as a member of jmj subfamily,JARID2 contains similar JmjC domain to JHDM1,JHDM2 Aand JARID1,separately.However the effect of JARID2 the activity on H3K9me3andH3K27me3 methylation is still unknown.In this study,we inhibited mRNA expressing level of JARID2 using the method of RNAi and detected H3K4me3,H3K9me3,H3K27me3 and H3K36me3 methylation level by immunofluorescence technology.The results showed that JARID2 the expression of bovine cumulus cells,and the presence of H3K9me3,H3K27me3 methylation were detected,but without H3K4me3,H3K36me3 of methylation.Comparing to the control group,there was significantly decline of JARID2 expressing level in transfected groups after the mRNA expressing of JARID2 was inhibited successfully in bovine cumulus cells through RNAi.But H3K4me3,H3K27me3 expression were no significant changes,and H3K9me3,H3K27me3 expression decreased significantly,indicating that JARID2 at the mRNA level can affect the methylation degree of H3K9me3 and H3K27me3,but no effect for H3K4me3 and H3K36me3.