miR-221-3p靶向TIMP-2对腹主动脉瘤血管平滑肌细胞增殖、凋亡影响  

Effect of miR-221-3p targeting TIMP-2 on proliferation and apoptosis of vascular smooth muscle cells in abdominal aortic aneurysm

作　　者：赵伦德[1] 刘胜[1] 尹庆雨 金巍 刘晓 ZHAO Lun-de;LIU Sheng;YIN Qing-yu;JIN Wei;LIU Xiao(Department of General Surgery,the First Affiliated Hospital of Xinxiang Medical College,Xinxiang,Henan 453100;Department of General Surgery 1,the First Affiliated Hospital of Henan Polytechnic University(The Second People's Hospital of Jiaozuo),Jiaozuo,Henan 454003,China)

机构地区：[1]新乡医学院第一附属医院普外科,河南新乡453100 [2]河南理工大学第一附属医院(焦作市第二人民医院)普外一区,河南焦作454003

出　　处：《热带医学杂志》2021年第9期1119-1124,共6页Journal of Tropical Medicine

基　　金：河南省医学科技攻关计划联合共建立项项目(LHGJ20200833)。

摘　　要：目的分析miR-221-3p靶向基质金属蛋白酶抑制因子-2(TIMP-2)对腹主动脉瘤(AAA)血管平滑肌细胞(VSMC)增殖和凋亡的影响。方法对人VSMC细胞进行培养,10μmol/L血管紧张素(AngⅡ)对VSMC进行处理,将VSMC细胞分为miR-221-3p inhibitor组(转染miR-221-3p inhibitor)、阴性对照组(转染miR-221-3p inhibitor NC)、空白组(未经转染的VSMC细胞),对照组(未进行AngⅡ处理及转染的VSMC细胞),实时荧光定量聚合酶链式反应(qRT-PCR)检测各组细胞中miR-221-3p、TIMP-2 mRNA的表达水平;细胞计数试剂盒8(CCK-8)法检测各组细胞活力;流式细胞术检测各组细胞凋亡情况;蛋白印迹(WB)法检测增殖蛋白β-连环蛋白(β-catenin)、细胞周期蛋白D1(CyclinD1)、凋亡蛋白B细胞淋巴瘤因子2(Bcl-2)、Bcl-2相关蛋白(Bax)及蛋白TIMP-2、基质金属蛋白酶-2(MMP-2)、MMP-12的表达;双荧光素酶报告实验证明miR-221-3p与TIMP-2的靶向关系。结果与对照组比较,空白组、阴性对照组miR-221-3p表达、细胞凋亡率、蛋白Bax、MMP-2、MMP-12表达显著升高,TIMP-2 mRNA表达水平、细胞活力、蛋白β-catenin、CyclinD1、Bcl-2、TIMP-2表达显著降低,差异均有统计学意义(P<0.05);与空白组、阴性对照组比较,miR-221-3p inhibitor组miR-221-3p表达、细胞凋亡率、蛋白Bax、MMP-2、MMP-12表达显著降低,TIMP-2mRNA表达水平、细胞活力、蛋白β-catenin、CyclinD1、Bcl-2、TIMP-2表达显著升高,差异均有统计学意义(P<0.05)。miR-221-3p靶向调控TIMP-2。结论Mi R-221-3p可能靶向抑制TIMP-2的表达,抑制VSMC的增殖,促进其凋亡,进而参与AAA的发病。Objective To investigate the effects of miR-221-3p on the proliferation and apoptosis of vascular smooth muscle cells(VSMC)in abdominal aortic aneurysm(AAA)by targeting tissue inhibitor of metalloproteinase-2(TIMP-2).Methods Human VSMC were cultured and treated with 10μmol/L angiotensinⅡ(AngⅡ);VSMC was divided into miR-221-3p inhibitor group,NC group,blank group and control group;the expression levels of miR-221-3p and TIMP-2 mRNA were detected by real-time quantitative polymerase chain reaction(qRT-PCR);the cell viability was detected by cell counting kit-8(CCK-8)method and apoptosis was detected by flow cytometry;the expression of proliferation proteinβ-catenin,CyclinD1 apoptosis protein B-cell leukemia/lymphoma 2(Bcl-2),Bcl-2 assaciated X protein(Bax),TIMP-2,matrix metalloproteinase-2(MMP-2)and MMP-12 were detected by western blot(WB)method;the targeting relationship between miR-221-3p and TIMP-2 was proved by double luciferase assay.Results Compared with those in the control group,the expression of miR-221-3p,apoptosis rate,the expression of Bax,MMP-2 and MMP-12 were significantly increased in the blank group;the expression of TIMP-2 mRNA,cell viability,the expression levels ofβ-catenin,CyclinD1,Bcl-2 and TIMP-2 were significantly decreased(P<0.05).Compared with those in blank group and NC group,the expression of miR-221-3p,apoptosis rate,the expression of Bax,MMP-2 and MMP-12 were significantly decreased in the miR-221-3p inhibitor group;the expression of TIMP-2 mRNA,cell viability,the expression ofβ-catenin,CyclinD1,Bcl-2 and TIMP-2 were significantly increased(P<0.05).Conclusion Mi R-221-3p might targetingly inhibit the expression of TIMP-2,inhibit the proliferation of VSMC,promote its apoptosis,and participate in the pathogenesis of AAA.

关 键 词：miR-221-3p 基质金属蛋白酶抑制因子-2 血管平滑肌细胞 增殖 凋亡 

分 类 号：R735.5[医药卫生—肿瘤]