The effects of hypoxia on silencing HLA⁃G expression and biological behaviors of JEG⁃3 cells and EPAS1 / 安徽医科大学学报

ABSTRACT

Objective@#To investigate the effect of silencing human leukocyte⁃associated antigen⁃G ( HLA⁃G) expression in the chorionic trophoblast cell line JEG⁃3 cells under hypoxic conditions on the biological function of JEG⁃3 cells and through the hypoxia response pathway of endothelial PAS1 region protein 1 (EPAS1)is involved in the molecular mechanism of preeclampsia under high altitude hypoxia. @*Methods@#The expression of HLA⁃G in JEG⁃3 cells was inhibited by transfection of small interfering RNA (siRNA) . The JEG⁃3 cells after transfection were divided into four groups : normoxic control group , hypoxic control group , normoxic inhibition group , and hypoxic inhibition group. CCK⁃8 test and Transwell test were used to detect the proliferation and invasion ability of the cells in four groups ; The effects of four groups of apoptosis and cell cycle were detected by flow cytometry; HLA⁃G and EPAS1 mRNA and protein expression levels were detected by real⁃time fluorescence quantitative PCR ( qPCR) and Western blot.@*Results @#① Compared with the normoxic control group , hypoxic control group , normoxic inhibition group , and hypoxic inhibition group could reduce the proliferation activity and invasion ability of JEG⁃3 cells , and significantly increase the apoptosis rate. The hypoxic control group and hypoxic inhibition group also produced anobvious necrotic cell population ; Under the condition of hypoxia , after reducing the expression of HLA⁃G , the cell necrosis rate was further aggravated ; Whether under normoxia or hypoxia , inhibition of HLA⁃G expression caused the cells to be blocked in the G1 phase. ② Compared with the normoxic control group , hypoxia control group , normoxic inhibition group , and hypoxia inhibition group decreased the expression of HLA⁃G protein , and hypoxia and inhibition of HLA⁃G had a synergistic effect; Hypoxia⁃inducible factors⁃2α ( HIF⁃2α ) , vascular endothelial growth factor(VEGF) and endothelin⁃1(ET⁃1) protein expression could be added , inhibition of HLA⁃G decreased the expression of inducible nitric oxide synthase( NOS2) . @*Conclusion@#In the hypoxic environment , silencing HLA⁃G may affect the biological behavior of trophoblast through the EPAS1 hypoxic response pathway and participate in the occurrence and development of preeclampsia.