干扰miR-429通过下调HIF-1α/VEGF通路促进子宫内膜异位症异位子宫内膜基质细胞的增殖、迁移和侵袭  

miR-429 promotes proliferation,migration and invasion of ectopic endometrial stromal cells in endometriosis by regulating HIF-1α/VEGF pathway

作　　者：秦琪 李会 李志英[1] QIN Qi;LI Hui;LI Zhiying(Department of Obstetrics and Gynecology,Affiliated Renhe Hospital of China Three Gorges University,Yichang,Hubei 443000,China)

机构地区：[1]三峡大学附属仁和医院妇产科,湖北宜昌443000

出　　处：《中国优生与遗传杂志》2023年第3期580-585,共6页Chinese Journal of Birth Health & Heredity

摘　　要：目的明确微小RNA(microRNA,miRNA/miR)-429是否通过调节缺氧诱导因子(HIF-1α)/血管内皮生长因子(VEGF)通路来影响子宫内膜异位症子宫内膜基质细胞(ESCs)的增殖、迁移和侵袭。取三峡大学附属仁和医院子宫内膜异位症患者(n=37)标本,作为EM组,并将同期的良性妇科疾病患者(n=37)标本作为对照Con组。将ESCs分为5组:正常内膜组(NC组)、异位内膜细胞组(EM组)、anti-miR-NC组(转染anti-miR-NC)、anti-miR-429组(转染miR-429 inhibitors)、anti-miR-429+Over-HIF-1α组(转染miR-429和Over-HIF-1α过表达质粒)方法实时定量PCR(RT-PCR)和Western blot检测miR-429、HIF-1α和VEGF的表达,并分析miR-429与HIF-1α、VEGF mRNA相关性。运用细胞计数试剂盒8(CCK-8)法、划痕实验和Transwell法测定细胞增殖、迁移与侵袭,Western blot检测HIF-1α、VEGF、上皮钙黏蛋白(E-cadherin)、神经钙黏蛋白(N-cadherin)表达。结果EM组较Con组miR-429表达量,HIF-1αmRNA、蛋白及VEGF mRNA、蛋白水平均显著增加(P<0.05),且miR-429与HIF-1αmRNA及VEGF mRNA表达均呈显著正相关关系(P<0.05)。与NC组相比,EM组、anti-miR-NC组miR-429表达量、HIF-1α、VEGF、N-cadherin蛋白水平、24 h、48 h、72 h的细胞活力、划痕愈合率、侵袭细胞数显著升高(P<0.05),E-cadherin蛋白数显著下调(P<0.05);与anti-miR-NC组相比,anti-miR-429组miR-429表达量、HIF-1α、VEGF、N-cadherin蛋白水平、24 h、48 h、72 h的细胞活力、划痕愈合率、侵袭细胞数显著下降(P<0.05),E-cadherin蛋白数显著升高(P<0.05);与anti-miR-429组相比,anti-miR-429+Over-HIF-1α组以上指标与促进干扰miR-429表达趋势相反。结论干扰miR-429表达可以下调HIF-1α/VEGF通路活性,进而抑制子宫内膜异位症ESCs的增殖、迁移和侵袭。Objective To investigate whether microRNA(miRNA/miR)-429 affects the proliferation,migration and invasion of endometrial stromal cells in endometriosis by regulating the hypoxia-inducible factor(HIF-1α)/vascular endothelial growth factor(VEGF)pathway.Take samples from patients with endometriosis(n=37)in our hospital as EM group,and take samples from patients with benign gynecological diseases(n=37)in the same period as control Con group.ESCs are divided into five groups:Normal intima group(NC group),ectopic intima cell group(EM group),anti-Mir-NC group(transfected with anti-miR-NC),anti-Mir-429 group(miR-429 inhibitors),anti-miR-429+Over-HIF-1α组(transfected with miR-429 and Over-HIF-1α plasmid).Methods Real-time quantitative PCR(RT-PCR)and Western blot were used to detect the expression of miR-429,HIF-1α and VEGF,and the correlation between miR-429 and HIF-1α and VEGF mRNA was analyzed.Cell proliferation,migration and invasion were measured by cell counting kit 8(CCK-8),scratch test and Transwell assay.The expressions of HIF-1α,VEGF,E-cadherin and N-cadherin were detected by Western blot.Results The expression of miR-429,HIF-1α and VEG in EM group were significantly increased compared with those in Con group(P<0.05),and there was a significant positive correlation between miR-429 and HIF-1α mRNA and VEGF mRNA expression(P<0.05).Compared with the NC group,the expression of miR-429,the protein levels of HIF-1α,VEGF,N-cadherin,the cell viability at 24 h,48 h,72 h,the wound healing rate,and the number of invasive cells in the EM and anti-miR-NC groups were significantly increased(P<0.05).The protein number of E-cadherin was significantly down-regulated(P<0.05).Compared with the anti-miR-NC group,the expression of miR-429,the protein levels of HIF-1α,VEGF and N-cadherin,the cell viability at 24 h,48 h and 72 h,the wound healing rate and the number of invasive cells in the anti-miR-429 group were significantly decreased(P<0.05).The number of E-cadherin protein was significantly increased(P<0.05).Compared wi

关 键 词：子宫内膜异位症 子宫内膜基质细胞 miR-429 HIF-1α/VEGF通路 

分 类 号：R711.71[医药卫生—妇产科学]