RNA sequencing of endometrial stromal cells transfected with scrambled or siRHOB siRNA and induced decidualization

Endometrial decidualization refers to the proliferation and differentiation of ESCs into DSCs. During decidualization, ESCs undergo cytoskeletal rearrangement-mediated morphological changes and express decidualization markers, such as IGFBP1 and PRL. Rho GTPases are mainly involved in the regulation of a variety of cellular processes, including cytoskeleton remodeling-mediated morphological changes, motility, endocytosis and adhesion, as well as cell cycle progression, cell differentiation and apoptosis. The key roles of Rho proteins in cytoskeletal rearrangement and cell differentiation suggest that they may play a role in regulating the process of endometrial decidualization. In our study, RhoB was found to be the most significantly upregulated protein after the in vitro decidualization of human ESCs. The expression of RhoB was upregulated mainly by cAMP-CREB signaling and partly by progesterone signaling. Knockdown of RhoB in ESCs greatly influenced actin cytoskeletal rearrangement and cell morphology and decreased the expression of IGFBP1. SEMA3A, a downstream target gene of RhoB, could promote ESC decidualization by binding its receptor, plexinA4. More importantly, the expression levels of RhoB, SEMA3A, and plexinA4 were all decreased in decidua from patients with miscarriage. This suggests that abnormal RhoB/SEMA3A/plexinA4 signaling, resulting in deficient endometrial decidualization, may be related to the pathogenesis of miscarriage. Our study reveals the important role of RhoB/SEMA3A/plexinA4 signaling in regulating ESC decidualization, which might be used as a target for pregnancy-related diseases caused by abnormal endometrial decidualization in early pregnancy.