Expression of GRHL2 in the placenta of preeclampsia patients and its impacts on proliferation and invasion of trophoblast cells
Objective To investigate the expression of the grainyhead-like 2(GRHL2)gene in the placenta of preeclampsia(PE)patients and its impacts on the proliferation and invasion of trophoblast cells.Methods Placental tissues were collected from 32 PE patients who underwent cesarean section at Shaanxi Provincial People's Hospital from January to June 2022(PE group).Placental tissues were also collected from 32 normal pregnant women who underwent cesarean section during the same period at the same hospital(control group).Western blot was used to detect the expression of GRHL2 protein in placental tissues.Human chorionic trophoblast cells HTR-8/Svneo in the logarithmic growth phase were divided into the blank group,GRHL2 small interfering RNA negative control(si-NC)group,GRHL2 small interfering RNA(si-GRHL2)group,GRHL2 overexpression plasmid negative control(pcDNA)group,and GRHL2 overexpression plasmid(pcDNA-GRHL2)group.CCK-8 and colony formation assays were used to detect the OD450 value and colony formation rate of HTR-8/Svneo cells.Transwell assay was used to detect HTR-8/Svneo cell invasion.Western blot was used to detect the expression of GRHL2,proliferating cell nuclear antigen(PCNA),matrix metalloproteinase(MMP)-2,phosphorylated phosphatidylinositol 3-kinase(p-PI3K),and phosphorylated serine/threonine-protein kinase(p-Akt)proteins in HTR-8/Svneo cells.Results The expression level of GRHL2 protein in the placental tissues of the PE group was significantly higher than that in the control group(t=57.240,P＜0.05).Compared with the blank group and si-NC group,the expression of GRHL2 protein in HTR-8/Svneo cells of the si-GRHL2 group was decreased,while the OD450 value,colony formation rate,number of invasive cells,and the expression levels of PCNA,MMP-2,p-PI3K,and p-Akt proteins were increased(t value ranged from 5.477 to 24.595,P＜0.05).Compared with the blank group and pcDNA group,the expression of GRHL2 protein in HTR-8/Svneo cells of the pcDNA-GRHL2 group was increased,while the OD450 value,colony formation rate,number of invasive cells,and the expression levels of PCNA,MMP-2,p-PI3K,and p-Akt proteins were decreased(t values ranged from 9.295 to 24.239,P＜0.05).Conclusion GRHL2 is highly expressed in the placental tissues of PE patients.Silencing GRHL2 can promote the proliferation and invasion of HTR-8/Svneo cells,while overexpressing of GRHL2 can inhibit the proliferation and invasion of HTR-8/Svneo cells.The mechanism may be related to the regulation of the PI3K/Akt pathway.
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