The Function Of MiR-302/367 For Maintenance Of Self-renewal And Pluripotency In Mouse F9 Cells

Micro RNAs are the small noncoding RNAs,whose length is about 18-25 nt. They bind to the targets’ 3’UTR by base complementation pairing, and then guide the RNAinduced silence complex to target sites, which affects the mRNA stability and/or translational efficiency, as well as regulate cell biological progress. Due to the specific temporal and spatiae characters, they play important roles in many biological progresses, such as cell differentiation, cell proliferation, apoptosis and development. There is hardly any study about the functions of miR-302/367 for the maintence of self-renewal and pluripotency in mouse F9 cells. This study is aim to explore the functions of miR-302/367 for the maintence of self-renewal and pluripotency in mouse F9 cells through investageting how miR-302/367 affects the level of pluripotency factors, differentiation related factors and cell proliferation.The study found that the treatment of F9 cells with retinoic acid(RA), could chang cell morphology, and promote the expression of differentiation related factors such as aFP, Clnd6, Thbd and GATA6, while repress the pluripotency factors such as Klf4, Nanog. Alkaline phosphatase(AP) staining showed that miR-302/367 can maintain the cell morphology. In order to invetagte the effects of miR-302/367 on pluripotency factors, we use RT-qPCR and Western blot,and the results the results showed that miR-302/367 can promote the pluripotency factors’ mRNA and protein level, meanwhile it also can antagonize decrease of Nanog, Oct4 induced by RA, and recover the level of them partially. To test whether miR-302/367 regulate the differentiation related factors. By using the DLR assay, we found that Smad2 and Tgfbr2 were the targets of miR-302/367. In further measurement our results showed that miR-302/367 could repress the expression of Tgfbr2 and recover the Tgfbr2 to a normal level. Cell proliferation is very improtent to the maintence of self-renewal and pluripotency, to address the effect of miR-302/367 on cell proliferation, the CCK-8 cell number counting Kit was used and we found that miR-302/367 could promote the cell proliferation.In conclusion, miR-302/367 could matain self-renewal and pluripotency in mouse F9 cells via promotting the expression of pluripotency factors and the cell proliferation repressing the differentiation related factors.