Deciphering the role of PGRMC2 in the human endometrium during the menstrual cycle and in vitro decidualization using an in vitro approach

The endometrial response to progesterone is mediated by the classical and non-classical progesterone receptors. We previously demonstrated that PGRMC1 is critical for endometrial function, embryo implantation, and future placentation, however, the role(s) of PGRMC2, which is structurally similar to PGRMC1, have not been studied in the human endometrium. To reveal PGRMC2 function during the decidualization process, we specifically knocked down PGRMC2 with siRNAs in four hEnSC before in vitro decidualization. In vitro decidualization of hEnSCs was induced using co-treatment of cAMP and medroxyprogesterone 17-acetate progestin for four days, and evaluated by measuring prolactin by ELISA and F-actin immunostaining. After decidualization, we performed a RNA-seq of four PGRMC2 silencing d-hEnSC and four scramble d-hEnSC. The common differentially expressed genes (DEGs) and proteins (DEPs) were considered for downstream functional enrichment analysis.

子宫内膜对孕酮的应答由经典孕酮受体与非经典孕酮受体共同介导。我们此前的研究已证实，孕酮受体膜成分1（PGRMC1）对子宫内膜功能、胚胎着床及后续胎盘形成均至关重要；但结构与PGRMC1相似的孕酮受体膜成分2（PGRMC2）在人子宫内膜中的作用尚未得到研究。为揭示PGRMC2在蜕膜化过程中的功能，我们在体外诱导蜕膜化前，利用小干扰RNA（siRNAs）特异性敲低了4株人子宫内膜基质细胞（hEnSC）中的PGRMC2表达。采用环腺苷酸（cAMP）与醋酸甲羟孕酮（medroxyprogesterone 17-acetate progestin）联合处理4天，诱导hEnSCs发生体外蜕膜化；并通过酶联免疫吸附试验（ELISA）检测催乳素表达，结合纤维状肌动蛋白（F-actin）免疫染色对蜕膜化效果进行评估。蜕膜化完成后，我们对4株PGRMC2沉默的蜕膜化hEnSC以及4株转染阴性对照siRNA的蜕膜化hEnSC进行了RNA测序（RNA-seq）。后续将以两组共有的差异表达基因（DEGs）与差异表达蛋白（DEPs）为对象，开展下游功能富集分析。