低氧下沉默HLA-G对JEG-3细胞、EPAS1表达及其生物学行为的影响  

The effects of hypoxia on silencing HLA-G expression and biological behaviors of JEG-3 cells and EPAS1

作　　者：孙春蕾 杨娟[2] 谢莹莺 许文宇[1,2] 雷娟 刘朵朵 Sun Chunlei;Yang Juan;Xie Yingying;Xu Wenyu;Lei Juan;Liu Duoduo(Graduate School of Qinghai University,Xining 810000;Dept of Obstetrics,Affiliated Hospital of Qinghai University,Xining 810000)

机构地区：[1]青海大学研究生院,西宁810000 [2]青海大学附属医院产科,西宁810000

出　　处：《安徽医科大学学报》2023年第1期47-53,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81760276)。

摘　　要：目的探讨在低氧条件下,沉默绒毛膜滋养细胞系JEG-3细胞的人类白细胞相关抗原-G(HLA-G)表达对JEG-3细胞生物学功能的影响和通过内皮PAS1区域蛋白1(EPAS1)低氧反应通路参与高原低氧条件下子痫前期发病的分子机制。方法通过转染小干扰RNA(siRNA)抑制JEG-3细胞中HLA-G的表达,将转染后的JEG-3细胞分为4组:常氧对照组、低氧对照组、常氧抑制组、低氧抑制组。采用CCK-8实验及体外侵袭实验法检测4组细胞的增殖及侵袭能力;用流式细胞术检测4组细胞凋亡和细胞周期的影响;通过qPCR技术及Western blot法检测4组细胞中HLA-G和EPAS1 mRNA及蛋白的表达水平。结果(1)与常氧对照组相比,低氧对照组及常氧、低氧抑制组均可使JEG-3细胞的增殖活性、侵袭能力降低,使得细胞凋亡率显著升高,低氧对照组与低氧抑制组产生了明显的坏死细胞群。低氧条件下,降低HLA-G的表达会提高细胞坏死率。无论在常氧还是低氧下,抑制HLA-G表达均使细胞被阻滞在G1期。(2)与常氧对照组相比,低氧对照组及常氧、低氧抑制组均降低HLA-G蛋白表达,且低氧和抑制HLA-G蛋白表达二者有协同作用,均可增加缺氧诱导因子-2α(HIF-2α)、血管内皮生长因子(VEGF)及内皮素-1(ET-1)蛋白表达,抑制HLA-G则降低诱导型一氧化氮合成酶(NOS2)的表达。结论低氧环境下,沉默HLA-G可能通过EPAS1低氧反应通路影响滋养细胞生物学行为,参与了子痫前期的发生发展。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 an obvious 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 hypoxi

关 键 词：子痫前期 低氧 人类白细胞抗原G 内皮PAS1区域蛋白1 高海拔 

分 类 号：R714.252[医药卫生—妇产科学]