微小RNA-324-3p-PDRG1轴对血管瘤生物学行为的影响  

Effects of microRNA-324-3p-PDRG1 axis on the biological behavior of hemangioma

作　　者：江洋 何柏慧 刘振楠 史业弘 聂冉 李建[1] Jiang Yang;He Bohui;Liu Zhennan;Shi Yehong;Nie Ran;Li Jian(Department of Burn Plastic Surgery and Medical Aesthetics,First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,China)

机构地区：[1]新乡医学院第一附属医院烧伤整形与医疗美容科,卫辉453100

出　　处：《中华实验外科杂志》2024年第5期932-935,共4页Chinese Journal of Experimental Surgery

基　　金：2020年度河南省医学科技攻关计划项目(LHGJ20200512)。

摘　　要：目的:探讨miR-324-3p-PDRG1轴对血管瘤生物学行为的影响。方法:选取新乡医学院第一附属医院收治的20例血管瘤组织和癌旁组织作为研究对象,采用荧光定量聚合酶链反应(PCR)和蛋白质印迹法(Western blot)分析癌旁组织和肿瘤组织miR-324-3p和PDRG1的表达水平。采用荧光定量PCR分析人脐静脉内皮细胞(HUVEC)、人血管瘤内皮细胞HemECs和HDEC中miR-324-3p表达水平。将人血管瘤内皮细胞HemECs分为miRNA对照组和miR-324-3p组,采用细胞计数试剂盒(CCK-8)和克隆形成实验分析两组细胞的增殖能力;流式细胞术和原位缺口末端标记法(TUNEL)染色分析两组细胞的凋亡水平;采用划痕实验和Transwell实验分析两组细胞的迁移和侵袭能力,采用双荧光素酶报告基因技术分析miR-324-3p靶基因,采用蛋白质免疫印迹细胞靶基因的表达水平。组间计量数据比较采用t检验。结果:血管癌组织中miR-324-3p表达水平(0.98±0.08)明显低于癌旁组织表达水平(0.51±0.12),差异有统计学意义(t=14.410,P<0.05)。癌旁组织中PDRG1蛋白表达水平(0.98±0.16)明显低于血管瘤组织(1.81±0.24),差异有统计学意义(t=12.820,P<0.05)。人正常血管内皮细胞miR-324-3p表达水平(1.05±0.10)明显低于人血管瘤内皮细胞HemECs和HDEC(0.52±0.09、0.69±0.09),差异有统计学意义(t=9.768、6.662,P<0.05)。miRNA对照组细胞吸光度值和克隆形成率[(2.02±0.09)、(71.84±8.61)%]高于miR-324-3p组细胞[(1.44±0.11)、(44.31±6.38)%],差异有统计学意义(t=9.910、6.295,P<0.05)。miR-324-3p对细胞凋亡的影响:miRNA对照组细胞凋亡比例和TUENL阳性率[(3.44±1.07)%、(3.38±0.55)%]高于miR-324-3p组细胞[(19.48±2.62)%、(13.79±2.53)%],差异有统计学意义(t=13.840、9.831,P<0.05)。miRNA对照组细胞迁移和侵袭数量[(112.33±14.62)、(86.17±8.18)个]高于miR-324-3p组细胞[(79.83±7.57)、(44.67±10.73)个],差异有统计学意义(t=4.834、7.534,P<0.05)。PDRG1是miR-324-3p的靶�Objective To investigate the effects of microRNA(miR)-324-3p-PDRG1 axis on the biological behavior of hemangioma.Methods Hemangioma tissues and adjacent tissues from 20 patients in our hospital were selected as the research objects.The expression levels of miR-324-3p and PDRG1 in adjacent tissues and tumor tissues were analyzed by fluorescence quantitative polymerase chain reaction(PCR)and Western blotting.The expression level of miR-324-3p in human umbilical vein endothelial cells(HUVECs),human hemangioma endothelial cells HemECs and HDEC were analyzed by fluorescence quantitative PCR.The human hemangioma endothelial cells HemECs were divided into miRNA control group and miR-324-3p group.The proliferation ability of the two groups of cells was analyzed by cell counting kit-8(CCK-8)and colony formation experiments.The apoptosis level of the two groups of cells was analyzed by flow cytometry and terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)staining.The migration and invasion ability of the two groups of cells were analyzed by scratch experiment and Transwell experiment.The target gene of miR-324-3p was analyzed by dual luciferase reporter gene technology.The repayment level of the target gene of the cells was analyzed by Western blotting.The t test was used for comparison of measurement data between groups.Results The expression level of miR-324-3p in hemangioma tissues(0.98±0.08)was significantly lower than that in adjacent tissues(0.51±0.12,t=14.410,P<0.05).The protein expression level of PDRG1 in adjacent tissues(0.98±0.16)was significantly lower than that in hemangioma tissues(1.81±0.24,t=12.820,P<0.05).The expression level of miR-324-3p in normal human vascular endothelial cells(1.05±0.10)was significantly lower than that in human hemangioma endothelial cells HemECs and HDEC(0.52±0.09,0.69±0.09,t=9.768,6.662,P<0.05).The absorbance value and colony formation rate of cells in the miRNA control group[(2.02±0.09),(71.84±8.61)%]were significantly higher than those in the miR-324-

关 键 词：微小RNA 血管瘤 增殖 凋亡 迁移 

分 类 号：R732.2[医药卫生—肿瘤]