miR-518b靶向调控LAPTM4B在稽留流产发病中的作用  

Role of miR-518b targeting LAPTM4B in the pathogenesis of missed abortion

作　　者：张永艳 夏妍芬 林佳慧 李俏霞 植枝福[1] ZHANG Yongyan;XIA Yanfen;LIN Jiahui;LI Qiaoxia;ZHI Zhifu(Graduate School,Guangxi Medical University,Nanning 530021;Department of Gynaecology and Obstetrics,the Third People′s Hospital of Nanchang,Nanchang 330000)

机构地区：[1]广西医科大学研究生院,南宁530021 [2]南昌市第三人民医院妇产科,南昌330000

出　　处：《郑州大学学报（医学版）》2024年第4期496-501,共6页Journal of Zhengzhou University(Medical Sciences)

基　　金：广西自然科学基金项目(2017JJA10049);广西医科大学第一附属医院临床研究攀登计划项目(YYZS2020015)。

摘　　要：目的:探讨miR-518b在稽留流产发病中的作用。方法:①利用双荧光素酶报告实验验证miR-518b与LAPTM4B的靶向关系。②HTR8/SVneo细胞分别转染miR-518b mimic及其阴性对照(NC)质粒,24 h后实时荧光定量PCR法及Western blot法检测细胞中miR-518b、LAPTM4B mRNA及蛋白的表达。③HTR8/SVneo细胞分别转染miR-518b mimic NC、miR-518b mimic和miR-518b mimic+LAPTM4B过表达质粒24 h后,与人脐静脉血管内皮细胞混合,行成管实验。④收集22例稽留流产患者及24例同期正常妊娠并自愿行人工流产者(对照组)的绒毛组织,实时荧光定量PCR法检测miR-518b、LAPTM4B mRNA,免疫组化法检测微血管密度(MVD),Western blot法检测LAPTM4B蛋白的表达。结果:①双荧光素酶报告实验证实miR-518b与LAPTM4B存在靶向关系。②与miR-518b mimic NC组比较,miR-518b mimic组细胞中LAPTM4B蛋白和mRNA表达水平均下降(P<0.05)。③miR-518b mimic NC组、miR-518b mimic组及miR-518b mimic+LAPTM4B过表达质粒组新生小管数分别为(31.00±1.00)、(16.33±2.08)、(32.67±2.08),3组比较,差异有统计学意义(P<0.001);miR-518b mimic组小于NC组和miR-518b mimic+LAPTM4B过表达质粒组(P<0.05)。④稽留流产组绒毛组织中miR-518b表达高于对照组,LAPTM4B mRNA、蛋白表达水平及MVD均低于对照组(P<0.05)。稽留流产组miR-518b与LAPTM4B mRNA的表达、MVD呈负相关(r=-0.546,-0.439,P<0.001);LAPTM4B蛋白表达与MVD呈正相关(r=0.686,P<0.001)。结论:miR-518b可能通过调控LAPTM4B的表达,抑制绒毛滋养层细胞血管成管能力,参与稽留流产的发病。Aim:To investigate the role of miR-518b in the pathogenesis of missed abortion.Methods:The targeting binding relationship between miR-518b and LAPTM4B was validated using a dual luciferase assay.HTR8/SVneo cells were allocated into 2 groups,and transfected with miR-518b mimic and its negative control(NC)plasmid for 24 hours,respectively,and the expressions of LAPTM4B mRNA and protein were detected by RT-qPCR and Western blot.HTR8/SVneo cells were allocated into 3 groups,transfected with miR-518b mimic,NC,and miR-518b mimic+LAPTM4B overexpression plasmid for 24 hours,respectively,and then mixed with HUVEC for tube formation assay.Villous tissue from missed abortion group(22 cases)and normal early pregnancy group(24 cases,accepting abortion)was collected to detect the expressions of miR-518b,LAPTM4B mRNA by RT-qPCR,microvessel density(MVD)by immunohistochemistry,and LAPTM4B protein by Western blot.ResuIts:Dual luciferase assay result showed the targeting relationship between miR-518b and LAPTM4B.The expression levels of LAPTM4B protein and mRNA in miR-518b mimic group were lower than those in miR-518b mimic NC group(P<0.05).The number of tube for mations of miR-518b mimic NC group,miR-518b mimic group and miR-518b mimic+LAPTM4B overexpression group,was(31.00±1.00),(16.33±2.08),(32.67±2.08),the differences among the 3 groups were significant(P<0.001),and that of miR-518b mimic group was lower than those of the other 2 groups(P<0.05).The expression of miR-518b in villous tissue of the missed abortion group was higher than that of control group(P<0.05),while the LAPTM4B mRNA and protein expression,MVD were lower than those of control group(P<0.05).In the missed abortion group,the expression of miR-518b was negtively correlated with LAPTM4B mRNA expression and MVD(r=-0.546,-0.439,P<0.001);the expression of LAPTM4B protein was positively correlated with MVD(r=0.686,P<0.001).ConcIusion:miR-518b may inhibit the tubular formation of trophoblast cells by regulating the expression level of LAPTM4B,thereby contributing to

关 键 词：稽留流产 绒毛组织 血管形成 miR-518b LAPTM4B 

分 类 号：R714.21[医药卫生—妇产科学]