lncRNA HOTAIR调节miR-326/NUS1轴对宫腔粘连子宫内膜基质细胞纤维化的影响  

Influence of lncRNA HOTAIR on fibrosis of endometrial stromal cells in intrauterine adhesions by regulating miR-326/NUS1 axis

作　　者：陈国斌[1] 祝文峰 史文娟[1] 董倩 CHEN Guo-bin;ZHU Wen-feng;SHI Wen-juan(Department of Obstetrics and Gynecology,Shenzhen Maternity and Child Health Hospital,Shenzhen 518000,China)

机构地区：[1]深圳市妇幼保健院妇产科,518000

出　　处：《中国实用医药》2023年第1期165-172,共8页China Practical Medicine

基　　金：LncRNA-ATB通过吸附miR-200c调控FOXF2和Smad6表达参与宫腔粘连纤维化发病的研究(项目编号:A2022374)。

摘　　要：目的探讨长链非编码RNA(lncRNA)同源框转录反义RNA(HOTAIR)调控miR-326/NUS1对宫腔粘连(IUA)子宫内膜基质细胞纤维化的影响。方法收集首次确诊为IUA的18例患者的子宫内膜组织,另取同期因不孕症接受宫腔镜子宫内膜活检的18例患者的子宫内膜组织作为对照。采用实时荧光定量聚合酶链式反应(qRT-PCR)、western blot分别检测IUA子宫内膜组织、正常子宫内膜组织、人子宫内膜间质细胞(HESC细胞)、IUA HES细胞[转化生长因子-β1(TGF-β1)处理HESC细胞48 h]中HOTAIR、miR-326及NUS1蛋白表达;将IUA HESC细胞分组为Ct组、pcDNA组、pcDNA-HOTAIR组、si-NC组、si-HOTAIR组、si-HOTAIR+inhibitor NC组、si-HOTAIR+miR-326 inhibitor组,采用q RTPCR检测组织和细胞中HOTAIR、miR-326表达;CCK-8法检测IUA HESC细胞增殖;流式细胞术检测IUA HESC细胞凋亡;western blot检测组织和细胞中NUS1、Ⅰ型胶原蛋白α1链(COL1A1)、纤连蛋白(FN)、α-平滑肌肌动蛋白(α-SMA)蛋白表达;双荧光素酶报告基因实验验证HOTAIR与miR-326、miR-326与NUS1的关系;RNA pull down实验验证HOTAIR与miR-326的关系。结果IUA子宫内膜组织中HOTAIR、NUS1/甘油醛-3-磷酸脱氢酶(GAPDH)表达水平高于正常子宫内膜组织,miR-326表达水平低于正常子宫内膜组织,差异具有统计学意义(P<0.05)。IUA HESC细胞中HOTAIR、NUS1/GAPDH表达水平高于HESC细胞,miR-326表达水平低于HESC细胞,差异具有统计学意义(P<0.05)。pcDNA-HOTAIR组HOTAIR、NUS1/GAPDH表达水平高于Ct组、pcDNA组,miR-326表达水平低于Ct组、pcDNA组,差异具有统计学意义(P<0.05);si-HOTAIR组HOTAIR、NUS1/GAPDH表达水平低于Ct组、si-NC组,miR-326表达水平高于Ct组、si-NC组,差异具有统计学意义(P<0.05);si-HOTAIR组、si-HOTAIR+inhibitor NC组及si-HOTAIR+miR-326 inhibitor组HOTAIR表达水平比较,差异无统计学意义(P>0.05);si-HOTAIR+miR-326 inhibitor组miR-326表达水平低于si-HOTAIR组、si-HOTAIR+inhibitor NC组,NUS1/GAPDH表�Objective Endometrial tissues were collected from 18 patients who were first diagnosed with IUA, and endometrial tissues from 18 patients who underwent hysteroscopic endometrial biopsy for infertility during the same period were taken as controls. To discuss the influence of long non-coding RNA(lncRNA) HOX transcript antisense RNA(HOTAIR) on fibrosis of endometrial stromal cells in intrauterine adhesions(IUA) by regulating miR-326/NUS1 axis. Methods Endometrial tissues were collected from 18 patients who were first diagnosed with IUA, and endometrial tissues from 18 patients who underwent hysteroscopic endometrial biopsy for infertility during the same period were taken as controls. Quantitative real-time polymerase chain reaction(qRT-PCR) and western blot were used to detect the expression of HOTAIR, miR-326 and NUS1 protein in IUA endometrial tissue, normal endometrial tissue, human endometrial stromal(HESC) cells, and IUA HES cells [transforming growth factor-β1(TGF-β1) treated HESC cells for 48 h]. IUA HESC cells were separated into Ct group, pcDNA group, pcDNA-HOTAIR group, si-NC group, si-HOTAIR group, si-HOTAIR+inhibitor NC group,and si-HOTAIR+miR-326 inhibitor group. The expression of HOTAIR and miR-326 in tissues and cells was detected by qRT-PCR;the proliferation of IUA HESC cells was detected by CCK-8 method;the apoptosis of IUA HESC cells was detected by flow cytometry;western blot was applied to detect the expression of NUS1, collagen type I α1 chain(COL1A1), fibronectin(FN), and α-smooth muscle actin(α-SMA) proteins in tissues and cells;dual-luciferase reporter gene assay was used to verify the relationship between HOTAIR and miR-326, miR-326and NUS1;RNA pull down experiments were applied to verify the relationship between HOTAIR and miR-326.Results The expression levels of HOTAIR and NUS1/glyceraldehyde-3-phosphate dehydrogenase(GAPDH) in IUA endometrial tissues were higher than those in normal endometrial tissues, and the expression level of miR-326 was lower than that in normal endometrial t

关 键 词：同源框转录反义RNA miR-326 宫腔粘连 增殖 纤维化 

分 类 号：R711.74[医药卫生—妇产科学]