敲降同源异型盒C8对7因子诱导的体细胞重编程的抑制作用

Inhibitory effects of knocking down homeobox C8 on seven factors-induced somatic cell reprogramming

为探究7因子(Jdp2-Jhdm1b-Mkk6-Glis1-Nanog-Esrrb-Sall4)诱导的体细胞重编程机制,通过正向与反向遗传学方法、定量分析重编程关键分子事件及同时敲降同源异型盒C8(homeobox C8, Hoxc8)和SMAD家族成员6(SMAD family member 6, Smad6)等方式解析Hoxc8在细胞多能性网络重建过程中的相关作用。结果表明:敲降Hoxc8可显著抑制7因子诱导的体细胞重编程,但过表达Hoxc8对其无影响。敲降Hoxc8既不影响重编程中多能性标记基因的上调与体细胞标记基因的下调,也不影响间质-上皮转化(mesenchymal-epithelial transition, MET)标记基因与细胞增殖标记基因的表达。同时敲降Hoxc8与Smad6可使单独敲降Hoxc8导致的重编程效率降低情况得到恢复。综上所述,Hoxc8在7因子诱导的体细胞重编程中具有重要作用,为进一步揭示Hoxc8调控细胞命运转变的机制提供了参考。

To explore the mechanism of seven factors(Jdp2-Jhdm1b-Mkk6-Glis1-Nanog-Esrrb-Sall4)-induced somatic cell reprogramming, we analyzed the related role of homeobox C8(Hoxc8) in the process of pluripotency network reconstruction through forward and reverse genetics, quantitative analysis of the classic reprogramming process, and simultaneous knockdown of Hoxc8 and SMAD family member 6(Smad6). The results showed that knockdown of Hoxc8 could significantly inhibit seven factors-induced somatic cell reprogramming, but overexpression of Hoxc8 had no effects. Furthermore, knockdown of Hoxc8 neither impeded the up-regulation of pluripotency marker genes and down-regulation of somatic cell marker genes nor impeded the expressions of mesenchymal-epithelial transition(MET) marker genes and cell proliferation marker genes. It was also found that simultaneous knockdown of Hoxc8 and Smad6 could rescue the inhibitory effects caused by knocking down Hoxc8 alone. In conclusion, these results suggest that Hoxc8 plays a pivotal role in somatic cell reprogramming,which provides a reference for further revealing the mechanism of Hoxc8 regulating cell fate transition.