GRHL2在子癇前期患者胎盤中的表達及對滋養層細胞增殖、侵襲的影響

［摘 要］目的 探討粒狀頭樣2基因（GRHL2）在子癇前期（PE）患者胎盤中的表達及對滋養層細胞增殖、侵襲的影響。方法 收集2022年1月至6月期間在陜西省人民醫院進行剖宮產分娩的32例PE患者的胎盤組織（PE組），同時收集同期在本院進行剖宮產分娩的32例正常妊娠產婦的胎盤組織（對照組），Western blot檢測胎盤組織中GRHL2蛋白表達；將對數生長期的人絨毛膜滋養層細胞HTR-8/Svneo分為空白組、GRHL2小干擾RNA陰性對照（si-NC）組、GRHL2小干擾RNA（si-GRHL2）組、GRHL2過表達質粒陰性對照（pcDNA）組、GRHL2過表達質粒（pcDNA-GRHL2）組，CCK-8、克隆形成實驗檢測HTR-8/Svneo細胞OD450值、克隆形成率；Transwell實驗檢測HTR-8/Svneo細胞侵襲；Western blot檢測HTR-8/Svneo細胞中GRHL2、增殖細胞核抗原（PCNA）、基質金屬蛋白酶（MMP）-2、磷酸化磷脂酰肌醇3激酶（p-PI3K）、磷酸化絲氨酸蘇氨酸蛋白激酶（p-Akt）蛋白表達。結果 PE組患者胎盤組織中GRHL2蛋白表達水平高于對照組，差異有統計學意義（t=57.240，P＜0.05）；與空白組、si-NC組比較，si-GRHL2組HTR-8/Svneo細胞中GRHL2蛋白表達降低，OD450值、克隆形成率、侵襲細胞數、PCNA、MMP-2、p-PI3K、p-Akt蛋白表達升高（t值介于5.477～24.595之間，P＜0.05）；與空白組、pcDNA組比較，pcDNA-GRHL2組HTR-8/Svneo細胞中GRHL2蛋白表達升高，OD450值、克隆形成率、侵襲細胞數、PCNA、MMP-2、p-PI3K、p-Akt蛋白表達降低（t值介于9.295～24.239之間，P＜0.05）。結論 GRHL2在PE患者胎盤組織中高表達，沉默GRHL2可促進HTR-8/Svneo細胞增殖與侵襲，而過表達GRHL2可抑制HTR-8/Svneo細胞增殖與侵襲，其機制可能與調控PI3K/Akt通路有關。

［關鍵詞］粒狀頭樣2基因；子癇前期；胎盤；增殖；侵襲

Doi：10.3969/j.issn.1673-5293.2024.09.002

［中圖分類號］R173""" ［文獻標識碼］A

［文章編號］1673-5293（2024）09-0007-07

Expression of GRHL2 in the placenta of preeclampsia patients and its impacts

on proliferation and invasion of trophoblast cells

［Abstract］ 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 Peoples 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.

［Key words］ grainyhead-like 2;preeclampsia;placenta;proliferation;invasion

子癇前期（preeclampsia，PE）是一種伴有高血壓和蛋白尿的多系統血管綜合征，通常在妊娠20周后可觀察到［1-2］。全世界PE的發病率約為2%～8%，它是孕婦及其胎兒死亡和發病的主要原因［3］。盡管醫學取得了重大進展，但娩出胎兒和胎盤仍然是唯一已知的治療PE的方法［4］。……