STOX1在子痫前期滋养细胞炎症反应中的作用研究

Study on the role of STOX1 in the inflammatory of trophoblast cells in preeclampsia

目的探讨STOX1(storkhead box1)基因在人绒毛膜癌细胞系JEG3细胞模拟的滋养细胞炎症反应中的作用。方法采用人绒毛膜癌细胞系JEG3细胞株模拟滋养细胞,构建过表达/敲除STOX1基因稳转细胞系模型,实验组及对照组加入0.05 mmol/L阿司匹林,采用Transwell测定细胞迁移能力,并采用流式细胞仪检测各组细胞凋亡,实时定量PCR和蛋白质免疫印迹法检测细胞相关炎性、缺氧、凋亡因子的表达、生物学行为、分子生物学的m RNA及蛋白的相对表达量。结果过表达STOX1滋养细胞组中缺氧诱导因子α(HIF-1α)、内皮型一氧化氮合酶(e NOS)、诱导型一氧化氮合酶(i NOS)B淋巴细胞瘤-2(Bcl-2)均显著降低(P<0.01);同时,炎性反应相关因子核因子κB(NF-κB)、肿瘤坏死因子α(TNF-α)、白介素8(IL-8)及凋亡相关因子、半胱氨酸蛋白酶-3(Caspase-3)在基因及蛋白水平表达均明显升高(P<0.05)。敲除STOX1后HIF-1α(P<0.01)、e NOS(P<0.05)、i NOS(P<0.001)、抗凋亡基因Bcl-2(P<0.01)表达升高,而炎性反应NF-κB(P<0.05)、TNF-α(P<0.01)、IL-8(P<0.01)及Caspase-3(P>0.05)水平均表达降低。结论STOX1诱导JEG3细胞系炎性反应、促进缺氧及凋亡相关基因的表达,阿司匹林可能抑制STOX1的作用,减少JEG3细胞凋亡,减少炎症反应,改善滋养细胞缺氧,敲除STOX1与使用阿司匹林具有协同的作用。

Objective To investigate the STOX1(Storkhead box1)in the inflammatory response of trophoblast cells simulated by human choriocarcinoma cell line JEG3 cells.Methods The human choriocarcinoma cell line JEG3 cell line was used to mimic trophoblast cells,and a stable transitional cell line model with overexpression/knockdown of STOX1 gene was constructed.0.05 mmol/L aspirin was added to the experimental and control groups,respectively.Cell migration ability was measured by Transwell,and apoptosis was detected in each group by flow cytometry,and realtime quantitative PCR and protein immunoblotting were used to detect the expression of cell-associated inflammatory,hypoxic and apoptotic factors,biological behavior,and the relative expression of mRNA and protein of molecular biology.Results Hypoxia-inducible factorα(HIF-1α),endothelial-type nitric oxide synthase(eNOS),and inducible nitric oxide synthase(iNOS),and apoptosis B lymphocytoma-2(Bcl-2)were significantly reduced in the overexpression STOX1 trophoblast group(P<0.01);Meanwhile,inflammatory response-related factor nuclear factorκB(NF-κB),tumor necrosis factorα(TNF-α),interleukin 8(IL-8)and cysteine protease-3(Caspase-3)were significantly increased at both gene and protein levels(P<0.05).The expression of hypoxia-inducible genes HIF-1α(P<0.01),eNOS(P<0.05),and i NOS(P<0.001)and anti-apoptotic genes Bcl-2(P<0.01)was elevated after knockdown of STOX1,while inflammatory responses NF-κB(P<0.05),TNF-α(P<0.01),IL-8(P<0.01),and Caspase-3(P>0.05)levels were reduced.Conclusion STOX1 induces inflammatory response,promotes hypoxia and apoptosis-related gene expression in JEG3 cell line,aspirin may inhibit the effect of STOX1,reduce apoptosis in JEG3 cells,decrease inflammatory response and improve hypoxia in trophoblast cells,and knockdown of STOX1 has synergistic effect with the use of aspirin.