摘要
目的探讨粒状头样2基因(GRHL2)在子痫前期(PE)患者胎盘中的表达及对滋养层细胞增殖、侵袭的影响。方法收集2022年1月至6月期间在陕西省人民医院进行剖宫产分娩的32例PE患者的胎盘组织(PE组),同时收集同期在本院进行剖宫产分娩的32例正常妊娠产妇的胎盘组织(对照组),Western blot检测胎盘组织中GRHL2蛋白表达;将对数生长期的人绒毛膜滋养层细胞HTR-8/Svneo分为空白组、GRHL2小干扰RNA阴性对照(si-NC)组、GRHL2小干扰RNA(si-GRHL2)组、GRHL2过表达质粒阴性对照(pcDNA)组、GRHL2过表达质粒(pcDNA-GRHL2)组,CCK-8、克隆形成实验检测HTR-8/Svneo细胞OD_(450)值、克隆形成率;Transwell实验检测HTR-8/Svneo细胞侵袭;Western blot检测HTR-8/Svneo细胞中GRHL2、增殖细胞核抗原(PCNA)、基质金属蛋白酶(MMP)-2、磷酸化磷脂酰肌醇3激酶(p-PI3K)、磷酸化丝氨酸苏氨酸蛋白激酶(p-Akt)蛋白表达。结果PE组患者胎盘组织中GRHL2蛋白表达水平高于对照组,差异有统计学意义(t=57.240,P<0.05);与空白组、si-NC组比较,si-GRHL2组HTR-8/Svneo细胞中GRHL2蛋白表达降低,OD_(450)值、克隆形成率、侵袭细胞数、PCNA、MMP-2、p-PI3K、p-Akt蛋白表达升高(t值介于5.477~24.595之间,P<0.05);与空白组、pcDNA组比较,pcDNA-GRHL2组HTR-8/Svneo细胞中GRHL2蛋白表达升高,OD_(450)值、克隆形成率、侵袭细胞数、PCNA、MMP-2、p-PI3K、p-Akt蛋白表达降低(t值介于9.295~24.239之间,P<0.05)。结论GRHL2在PE患者胎盘组织中高表达,沉默GRHL2可促进HTR-8/Svneo细胞增殖与侵袭,而过表达GRHL2可抑制HTR-8/Svneo细胞增殖与侵袭,其机制可能与调控PI3K/Akt通路有关。
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 OD_(450) 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 OD_(450) 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 OD_(450) 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.
作者
张雅
刘晓红
ZHANG Ya;LIU Xiaohong(Department of Obstetrics,Shaanxi Provincial People's Hospital,Shaanxi Xi′an 710068,China)
出处
《中国妇幼健康研究》
2024年第9期7-13,共7页
Chinese Journal of Woman and Child Health Research
基金
陕西省重点研发计划项目(2020SF-037)。
关键词
粒状头样2基因
子痫前期
胎盘
增殖
侵袭
grainyhead-like 2
preeclampsia
placenta
proliferation
invasion
