The regulation study of miRNA-449a on LGR4 and endometrial stromal cells in goat

摘要

MicroRNAs (miRNA) are small, evolutionarily conserved noncoding RNA molecules that actrnas posttranslational regulators of gene expression in a sequence-specific manner, and miR-449rninduced cell differentiation or apoptosis in a mutually exclusive fashion. LGR4 regulate cellrngrowth, proliferation, and differentiation, and was thought to be involved in reproductivernsystems. Our previous Illumina Solexa sequencing results indicated a high expression of miR-rn449a in receptive endometrium compared with of pre-receptive endometrium in goat. And wernpredicated and experimentally validated LGR4 as a target of miR-449a. To explore the role ofrnmiR-449a in the development of receptive endometrium, miRNA mimic and inhibitor werernused to perform miR-449a overexpression or knockdown, respectively. Our results showedrnthat overexpression of miR-449a could dramatically reduce the mRNA and protein expressionrnlevels of LGR4 in caprine endometrial stromal cells (ESC) both at 24h and 48h, and miR-rn449a knockdown significantly increased the LGR4 mRNA at 24h but not 48h. Thernoverexpression of miR-449a also led to a notable apoptosis but not affect cell cycle. Inrnconclusion, we verified miR-449a down-regulated LGR4 in ESC and induced ESC apoptosisrnthough the extrinsic (FAS-mediated) pathway.