miR-517在子痫前期患者胎盘外泌体中的表达及其对滋养层细胞侵袭与T细胞增殖的影响  

Expression of miR-517 in placental exosomes from patients with preeclampsia and its effect on trophoblast cell invasion and T cell proliferation

作　　者：罗书[1] 李书明 黄勇[3] 关红琼[1] LUO Shu;LI Shuming;HUANG Yong;GUAN Hongqiong(Department of Obstetrics,the Second Affiliated Hospital of Hainan Medical College,Haikou,Hainan 570311,China;the Second Clinical Medical College of Hainan Medical College,Haikou,Hainan 570311,China;Department of Reproductive Medicine,the Second Affiliated Hospital of Hainan Medical College,Haikou,Hainan 570311,China)

机构地区：[1]海南医学院第二附属医院产科,海南海口570311 [2]海南医学院第二临床医学院,海南海口570311 [3]海南医学院第二附属医院生殖医学科,海南海口570311

出　　处：《国际检验医学杂志》2023年第12期1452-1458,共7页International Journal of Laboratory Medicine

基　　金：海南省卫生计生行业科研项目(20A200264)。

摘　　要：目的 探讨在子痫前期(PE)患者的胎盘组织与胎盘外泌体中微小RNA-517(miR-517)的表达及其对滋养层细胞侵袭与T细胞增殖的影响。方法 收集2020年5月至2021年5月于海南医学院第二附属医院确诊为PE的患者50例(PE组)和50例健康孕产妇(NP组)的胎盘绒毛组织和外周血,采用实时荧光定量PCR(qPCR)检测两组胎盘绒毛组织中miR-517的表达差异;分离外周血中的胎盘外泌体,采用透射电子显微镜(TEM)与纳米示踪技术(NTA)分析外泌体形态、大小分布及浓度,蛋白质印记法(Western blot)检测外泌体标志性蛋白CD63、肿瘤易感基101(TSG101)和胎盘特异性蛋白胎盘碱性磷酸酶(PLAP)的表达;体外培养人绒毛滋养层细胞HTR-8与人T细胞系Jurkat T细胞,将过表达miR-517的miR-517 mimic与阴性对照序列miR-NC转染至上述细胞中,并按细胞处理方式的不同将HTR-8与Jurkat T细胞各分为5组:对照组、miR-517 mimic组、miR-NC组、NP-外泌体组、PE-外泌体组;qPCR检测各组HTR-8或Jurkat T细胞中miR-517的表达水平,CCK-8检测其增殖能力;Transwell检测各组HTR-8细胞的侵袭能力。结果 与NP组比较,PE组胎盘组织与胎盘外泌体中miR-517的表达均明显降低(P<0.05),且胎盘外泌体具有典型的圆盘状或新月形的双层膜结构,颗粒直径在30~150 nm, CD63、TSG101、PLAP表达升高(P<0.05);与对照组比较,miR-517 mimic组与NP-外泌体组中miR-517表达和HTR-8细胞增殖及侵袭能力均明显升高(P<0.05),Jurkat T细胞的增殖能力却明显降低(P<0.05),miR-NC组均无明显改变(P>0.05),PE-外泌体组中miR-517水平无明显差异(P>0.05),但HTR-8细胞增殖及侵袭能力均降低(P<0.05),而Jurkat T细胞的增殖能力却明显升高(P<0.05)。与NP-外泌体组比较,miR-517 mimic组中miR-517表达和HTR-8细胞增殖、侵袭能力均明显升高(P<0.05),而Jurkat T细胞的增殖能力却降低(P<0.05),PE-外泌体组中miR-517表达和HTR-8细胞增殖、侵袭能力均明显降低(P<0.05),�Objective To investigate the expression of microRNA-517(miR-517)in placental tissues and placental exosomes of patients with preeclampsia(PE)and its effect on trophoblast cell invasion and T cell proliferation.Methods Placental villus tissue and peripheral blood were collected from 50 patients diagnosed with PE(PE group)and 50 healthy pregnant women(NP group)in the Second Affiliated Hospital of Hainan Medical College from May 2020 to May 2021.Real-time fluorescence quantitative PCR(qPCR)was used to detect the expression of miR-517 in placental villus tissue of the two groups.Placental exosomes were isolated from peripheral blood.Transmission electron microscopy(TEM)and nanotracer technology(NTA)were used to analyze the morphology,size distribution and concentration of exosomes.The expression of exosome marker cardiac protein CD63,tumor susceptibility gene 101(TSG101)and placental specific protein placental alkaline phosphatase(PLAP)were detected by Western blot.Human chorionic trophoblastic HTR-8 cells and human T cell line Jurkat T cells were cultured in vitro.miR-517 mimic and negative control sequence miR-NC were transfected into the above cells.HTR-8 and Jurkat T cells were divided into 5 groups according to different cell treatment methods:control group,miR-517 mimic group,miR-NC group,NP-exosome group and PE-exosome group,respectively.qPCR was used to detect the expression of miR-517 in HTR-8 or Jurkat T cells.CCK-8 was used to detect the proliferation ability of HTR-8 or Jurkat T cells,and Transwell was used to detect the invasion ability of HTR-8 cells in each group.Results Compared with the NP group,the expression of miR-517 in the placental tissue and placental exosomes in the PE group was significantly decreased(P<0.05).The placental exosomes in the PE group had a typical disc-shaped or crescent-shaped bilayer membrane structure,and the particle diameter was 30-150 nm,and the expression of CD63,TSG101 and PLAP proteins was increased(P<0.05).Compared with the control group,the expression of miR-517 and

关 键 词：子痫前期 微小RNA-517 胎盘外泌体 滋养层细胞 T细胞 增殖 侵袭 

分 类 号：R446.19[医药卫生—诊断学]