Study of the effect of cAMP signaling pathway on HTR8/SV-neo cell line proliferation,invasion and migration

Abstract

Abstract Pre-eclampsia (PE) is thought to be related to placental dysfunction, particularly poor extravillous trophoblast (EVT) invasion and migration abilities. However, the pathogenic mechanism is not yet fully understood. Here, we used the HTR8/SV-neo cell line as a model for the in vitro functional study of human extravillous trophoblasts. HTR8/SV-neo cells were treated with different concentrations of Forskolin solution (cAMP pathway-specific agonist) to alter intracellular cAMP level. Taking the DMSO group as the control group, firstly, cAMP assay kit was used to detect the cAMP concentration in HTR8/SV-neo cells treated with different concentration of Forskolin, and the cell proliferation function was detected by cell growth curve drawing and the colony formation experiment. The invasiveness and migration of cells were observed by Transwell experiments, and the expression of intracellular EMT-related markers was further evaluated by qPCR and WB experiments. According to our research, the intracellular cAMP level in HTR8/SV-neo cells was increased in a dose-dependently manner, and the proliferation, invasion and migration abilities of HTR8/SV-neo cells were significantly enhanced. Further detection by qPCR and WB experiments proved that the expression of EMT and angiogenesis markers in cells were up-regulated, including MMP2, PECAM, N-cadherin and VE-cadherin. In addition, with the increase of intracellular cAMP level, the phosphorylation level of intracellular MAPK signaling pathway was significantly increased. Thus, these results provide a novel rationale for the treatment of PE patients by improving the poor placental environment.