妊娠期糖尿病中细胞因子信号传导抑制蛋白-3过表达对滋养细胞增殖与凋亡的作用研究  

Effect of overexpression of suppressor of cytokine signaling-3 on trophoblast proliferation and apoptosis in gestational diabetes mellitus

作　　者：刘晶晶[1] 刘莹 李玉珍 LIU Jingjing;LIU Ying;LI Yuzhen(Department of Obstetrics and Gynecology,The Fourth Hospital of Xi'an;School of Basic Medicine,School of Medicine,Xi'an Jiaotong University,Xi'an Shaanxi 710075,P.R.China)

机构地区：[1]西安市第四医院妇产科,陕西西安710075 [2]西安交通大学医学部基础医学院,陕西西安710075

出　　处：《中国计划生育和妇产科》2021年第6期60-65,共6页Chinese Journal of Family Planning & Gynecotokology

基　　金：陕西省科技计划项目(项目编号:2019SF-164)。

摘　　要：目的探讨细胞因子信号传导抑制蛋白-3(suppressor of cytokine signaling-3,SOCS-3)在妊娠期糖尿病(gestational diabetes mellitus, GDM)患者胎盘组织中的表达,及其过表达对滋养细胞增殖和凋亡的影响。方法收集西安市第四医院2018年6月至2019年12月50例GDM患者和50例健康孕妇的胎盘组织,采用荧光定量PCR(qRT-PCR)检测胎盘组织中SOCS-3 mRNA的表达,免疫组织化学染色检测SOCS-3蛋白表达。将培养的HTR-8/SVneo细胞随机分为空白对照组、空质粒转染组与SOCS-3质粒转染组,采用Lipofectamine 2000将pCR 3.1/SOCS-3质粒载体和pCR 3.1空质粒载体分别转染至SOCS-3质粒转染组与空质粒转染组细胞中,qRT-PCR和蛋白质免疫印迹(Western blot)检测转染效率,并通过CCK-8、流式细胞术、Annexin V-FITC/PI及Hoechst 33342实验分别观察过表达SOCS-3对细胞活性、周期分布与凋亡的影响。结果 SOCS-3在GDM患者胎盘组织中表达量明显高于健康胎盘组织;体外转染pCR 3.1/SOCS-3质粒载体后,SOCS-3质粒转染组细胞中SOCS-3 mRNA及蛋白的表达量较空白对照组明显升高;与空白对照组相比,SOCS-3质粒转染组HTR-8/SVneo细胞活性下降,S期细胞比例显著升高而G1期细胞比例下降,细胞凋亡率增加,且出现明显的细胞凋亡形态。结论 SOCS-3在GDM患者胎盘组织中呈现高表达,过表达SOCS-3可显著抑制滋养细胞的增殖并促进其凋亡。Objective To investigate the expression of suppressor of cytokine signaling-3(SOCS-3)in placental tissue of patients with gestational diabetes mellitus(GDM),and the effect of over expression on trophoblast proliferation and apoptosis.Methods Collected placental tissues of 50 GDM patients and 50 healthy pregnant women from June 2018 to december 2019 in The Fourth Hospital of Xi’an.Fluorescence quantitative PCR(qRT-PCR)was used to detect the expression of SOCS-3 mRNA in placental tissue,immunohistochemical staining was used to detect SOCS-3 protein expression.The cultured HTR-8/SVneo cells were randomly divided into blank control group,empty plasmid transfection group and SOCS-3 plasmid transfection group,using Lipofectamine 2000,the pCR3.1/SOCS-3 plasmid vector and pCR3.1 empty plasmid vector were transfected into SOCS-3 plasmid transfection group and empty plasmid transfection group cells,respectively.qRT-PCR and Western blot were used to detect transfection efficiency.CCK-8,flow cytometry,Annexin V-FITC/PI and Hoechst 33342 experiments were used to observe the effects of overexpression of SOCS-3 on cell activity,cycle distribution and apoptosis.Results The expression of SOCS-3 in the placental tissue of GDM patients was significantly higher than that of healthy placenta tissue.After transfection of pCR3.1/SOCS-3 plasmid vector in vitro,the expression of SOCS-3 mRNA and protein in cells of SOCS-3 plasmid transfection group was significantly higher than that of blank control group.Compared with the blank control group,the activity of HTR-8/SVneo cells in the SOCS-3 plasmid transfection group decreased,the proportion of cells in S phase increased significantly and the proportion of cells in G1 phase decreased,apoptosis rate increased significantly,and there was an obvious form of apoptosis.Conclusion SOCS-3 is highly expressed in the placental tissue of GDM patients.Overexpression of SOCS-3 can significantly inhibit the proliferation of trophoblast cells and promote their apoptosis.

关 键 词：妊娠期糖尿病 细胞因子信号传导抑制蛋白-3 滋养细胞 增殖 凋亡 

分 类 号：R737.33[医药卫生—肿瘤]