miR-219、IGF2 mRNA在早期胎停中的表达及对绒毛滋养层细胞迁移、侵袭的影响  

The expression of miR-219 and IGF2 mRNA in early fetal arrest and their effects on the migration and invasion of trophoblast cells

作　　者：牛会坤 李博 韩晶[1] 吴娜 张会辰[1] 薛楠 NIU Huikun;LI Bo;HAN Jing;WU Na;ZHANG Huichen;XUE Nan(Baoding Maternal and Child Health Hospital,Baoding 071000,China)

机构地区：[1]河北省保定市妇幼保健院妇产科,071000

出　　处：《中国煤炭工业医学杂志》2024年第6期541-547,共7页Chinese Journal of Coal Industry Medicine

基　　金：河北省医学科学课题计划(编号:202002710)

摘　　要：目的 分析早期胚胎停止发育绒毛组织中微小RNA-219(miR-219)、胰岛素样生长因子2(IGF2) mRNA表达水平,探讨二者对胎盘绒毛滋养层细胞迁移和侵袭的影响。方法 随机选取2022年5月—2024年5月保定市妇幼保健院收治的早期胚胎停止发育的88例患者作为研究组,另外选取55例同期早孕人工流产者作为对照组,实时荧光定量PCR(qPCR)方法检测两组绒毛组织中miR-219、IGF2 mRNA表达。取购买的胎盘绒毛滋养层细胞系(HTR-8/SVneo),随机分为对照组、低氧组(使用100μmol/L氯化钴培养)、低氧+miR-219 mimics组(转染miR-219 mimics后使用100μmol/L氯化钴培养)、低氧+miR-219 mimics+oe-IGF2组(转染miR-219 mimics+IGF2过表达质粒后使用100μmol/L氯化钴培养),使用蛋白质印迹(WB)法检测各组基质金属蛋白酶2(MMP-2)、MMP-9蛋白表达量,四甲基偶氮唑盐比色(MTT)法、克隆形成实验检测HTR-8/SVneo细胞增殖情况,采用划痕试验检测细胞迁移情况,采用Transwell小室检测细胞侵袭能力。结果 人群研究显示:与对照组比较,研究组绒毛组织中miR-219表达较高,IGF2 mRNA表达较低(P<0.05)。细胞实验研究显示:与对照组比较,低氧组、低氧+mi R-219 mimics组、低氧+mi R-219 mimics+oe-IGF2组mi R-219表达量升高,IGF2 mRNA表达量、MMP-2、MMP-9蛋白表达量降低,增殖活性、克隆形成数、迁移、侵袭细胞数减少(P<0.05);与低氧组比较,低氧+miR-219 mimics组miR-219表达量升高,IGF2 mRNA表达量、MMP-2、MMP-9蛋白表达水平降低,增殖活性、克隆形成数、迁移、侵袭细胞数减少,低氧+miR-219 mimics+oe-IGF2组miR-219表达量降低,IGF2 mRNA表达量、MMP-2、MMP-9蛋白表达水平升高,增殖活性、克隆形成数、迁移、侵袭细胞数增加(P<0.05);与低氧+miR-219 mimics组比较,低氧+miR-219 mimics+oe-IGF2组miR-219表达量降低,IGF2 mRNA表达量、MMP-2、MMP-9蛋白表达水平升高,增殖活性、克隆形成数、迁移、侵袭细胞数增加(P<0.Objective Analyze the expression levels of microRNA-219(miR-219)and the mRNA expression of insulinlike growth factor 2(IGF2)in early embryonic trophoblast tissue,and explore their effects on the migration and invasion of placental trophoblast cells.Methods A randomized method was used to select 88 patients with early embryo arrest admitted to Baoding Maternal and Child Health Hospital from May 2022 to May 2024 as the study group,and 55 patients with early pregnancy induced abortion during the same period as the control group.Realtime quantitative PCR(qPCR)was used to detect the expression of miR-219 and IGF2 in villi of the two groups.Take the purchased placental trophoblast cell line(HTR-8/SVneo).They were divided into control group,hypoxia group(100μmol/L cobalt chloride culture),hypoxia+miR-219 mimics group(100μmol/L cobalt chloride culture after transfection of miR-219 mimics),hypoxia+miR-219 mimics+oe-IGF2 group(transfection of miR-219 mimics+IGF2 overexpressed plasmid was cultured with 100μmol/L cobalt chloride).Western Blot method was used to detect the protein of Matrix metalloproteinase(MMP-2)and MMP-9 in each group.The proliferation of HTR-8/SVneo cells was detected by MTT assay and clone formation experiment,the cell migration was detected by scratch test,and the cell invasion was detected by Transwell chamber.Results Compared with control group,the expression of miR-219 was higher and the expression of IGF2 was lower in villus tissues of the study group(P<0.05).Compared with the control group,the hypoxia group,hypoxia+miR-219 mimics group,and hypoxia+miR-219 mimics+oe-IGF2 group showed an increase in miR-219 expression,a decrease in IGF2 mRNA expression,matrix metalloproteinase-2(MMP-2),and MMP-9 protein expression,and a decrease in proliferation activity,clone formation,migration,and invasion cell numbers(P<0.05).Compared with the hypoxia group,the hypoxia+miR-219 mimics group showed an increase in miR-219 expression,a decrease in IGF2 mRNA expression,MMP-2 and MMP-9 protein expression,and a decre

关 键 词：早期胚胎停止发育 胎盘绒毛滋养层细胞 微小RNA-219 胰岛素样生长因子2 

分 类 号：R714.51[医药卫生—妇产科学]