Hypoxia-induced GPR4 suppresses trophoblast cell migration and proliferation through the MAPK signaling pathway

Pre-eclampsia (PE), a common pregnancy-systemic syndrome, is characterized by proteinuria and hypertension and is the leading cause of maternal and fetal mortality. Thus, we aim to investigate the role of G-Protein Coupled Receptor 4 (GPR4) in PE and the underlying molecular mechanism. In this study, GSE66273 microarray data were obtained from the Gene Expression Omnibus(GEO) database of the National Center for Biotechnology Information, and Gene set enrichment analysis (GSEA) was performed by GSEA software. qRT-PCR and Immunohistochemistry (IHC) or western blotting were used to assay for the expression of GPR4 in PE placentas and HTR8/SVneo cells. The influence of acidosis and hypoxia environments on the expression of GPR4 was explored using western blotting. Cell proliferation and migration of HTR8/SVneo cells were measured using EdU and MTT assays and migration assay, respectively. Moreover, expressions of MEK1/2, p-MEK1/2, ERK1/2, and p-ERK1/2 in HTR8/SVneo cells were assayed by western blotting. Our data demonstrated that the expression of GPR4 was up regulated in PE placentas. Increase in acidic pH and hypoxic levels increased the expression of GPR4 in HTR8/SVneo cells. GPR4 inhibited cell proliferation and migration in the HTR8/SVneo cells. GPR4 silencing enhanced the phosphorylation of p-MEK1/2and p-ERK1/2 in HTR8/SVneo cells. Additionally, we found that pathway inhibitor partially reversed the effects of GPR4 on proliferation and migration of HTR8/SVneo cells. In conclusions, these results show that GPR4 suppressed cell proliferation and migration by inhibiting MAPK signaling pathway in PE.