DNMT3B基因对猪睾丸支持细胞迁移及下游相关基因表达的影响

Effects of DNMT3B gene on migration and downstream related gene expression of porcine sertoli cells

睾丸支持细胞(sertoli cells,ST)是最重要的生殖相关细胞,对精子的发育具有重要的支持功能,DNA甲基转移酶3B(DNA methyltransferase 3 beta,DNMT3B)是参与表观遗传调控的重要基因之一,在猪的雄性遗传信息传递中具有重要的作用。为了探讨DNMT3B基因对猪睾丸支持细胞增值及下游相关基因表达的影响,试验通过在ST细胞系中过表达DNMT3B基因,借助细胞划痕试验检测细胞的迁移能力,通过荧光定量PCR方法检测其对部分组蛋白、热应激、凋亡相关基因表达的影响。结果表明:与对照组相比,过表达DNMT3B基因的猪ST细胞相对迁移速率受到抑制,在划痕后第24,48小时时抑制作用明显(P<0.05)。过表达DNMT3B基因后DNMT3A、HSF1、P53、C-myc相对表达量显著提高(P<0.05)。说明DNMT3B对特定的热应激和细胞凋亡基因有直接的影响,但未发现其对特定组蛋白相关基因有明显调节作用。

Sertoli cells(ST)are the most important reproduction-related cells and have an important supporting function for sperm development.DNA methyltransferase 3 beta(DNMT3B)is one of the important genes involved in epigenetic regulation and plays an important role in the transmission of male genetic information in pigs.To investigate the effect of DNMT3B gene on the proliferation of pig sertoli cells and the expression of downstream related genes,this study through a DNMT3B gene expression in the ST cell lines,with the help of cell of scratch test cell migration ability,RT-PCR method to detect the downstream and methylation,heat shock,apoptosis related gene expression changes.It is found that compared with control group,with the overexpression of DNMT3B gene ST pig cells the relative migration rate is restrained,when Nick after 24 h,48 h inhibition significantly(P<0.05).The relative expression levels of DNMT3A,HSF1,P53 and C-myc were significantly increased after DNMT3B overexpression(P<0.05).It is suggested that DNMT3B has a direct effect on specific heat stress and apoptosis genes,but no obvious regulation effect on specific histone related genes was found.