miR-519e-5p靶向ANXA5对脊索瘤U-CH1细胞生物学行为的影响  

Effect of miR-519e-5p targeting ANXA5 on the biological behavior of chordoma U-CH1 cells

作　　者：李轶 黄小平 王玮 LI Yi;HUANG Xiao-ping;WANG Wei(Department of Vascular Intervention,Xi'an Ninth Hospital,Xi'an Shaanxi 710054,China;Department of Gastroenterology,Xi'an Ninth Hospital,Xi'an Shaanxi 710054,China;Department of Oncology and Hematology,Xi'an Ninth Hospital,Xi'an Shaanxi 710054,China)

机构地区：[1]西安市第九医院血管介入科,陕西西安710054 [2]西安市第九医院消化内科,陕西西安710054 [3]西安市第九医院肿瘤血液科,陕西西安710054

出　　处：《局解手术学杂志》2024年第10期892-897,共6页Journal of Regional Anatomy and Operative Surgery

基　　金：西安市第九医院科研项目(2020-7)。

摘　　要：目的探讨miR-519e-5p靶向膜联蛋白A5(ANXA5)对脊索瘤U-CH1细胞生物学行为的影响。方法以qRT-PCR测定人脊索瘤组织和癌旁组织中miR-519e-5p和ANXA5的表达。体外培养U-CH1细胞,并分为miR-519e-5p mimics组、miR-519e-5p mimics阴性对照+空载组、miR-519e-5p mimics+ANXA5过表达组、对照组。平板集落形成实验、MTT实验检测各组U-CH1细胞增殖;流式细胞术检测各组U-CH1细胞凋亡;划痕实验、Transwell实验分别检测各组U-CH1细胞迁移与侵袭;Western blot检测各组U-CH1细胞ANXA5蛋白、凋亡相关蛋白(Bcl-2、Bax)及迁移上皮-间充质转化相关蛋白(Claudin-1、Vimentin、E-cadherin)表达;qRT-PCR法检测各组U-CH1细胞miR-519e-5p及ANXA5 mRNA表达;双荧光素酶报告基因实验检验miR-519e-5p与ANXA5在U-CH1细胞中的相互作用。结果与癌旁组织相比,脊索瘤组织miR-519e-5p表达降低(P<0.05),ANXA5 mRNA表达升高(P<0.05)。与对照组相比,miR-519e-5p mimics组细胞增殖率、集落生成率、迁移率、侵袭数、ANXA5 mRNA及ANXA5、Bcl-2、Claudin-1、Vimentin蛋白表达降低(P<0.05),miR-519e-5p mRNA表达、凋亡率及Bax、E-cadherin蛋白表达增高(P<0.05)。与miR-519e-5p mimics组相比,miR-519e-5p mimics+ANXA5过表达组细胞集落生成率、增殖率、侵袭数、迁移率、ANXA5 mRNA及ANXA5、Bcl-2、Vimentin、Claudin-1蛋白表达升高(P<0.05),凋亡率及Bax、E-cadherin蛋白表达下降(P<0.05)。miR-519e-5p可靶向下调U-CH1细胞中ANXA5表达。结论miR-519e-5p通过靶向抑制ANXA5表达降低脊索瘤U-CH1细胞迁移、增殖和侵袭活力,促进其凋亡。Objective To investigate the effect of miR-519e-5p targeting Annexin A5(ANXA5)on the biological behavior of chordoma U-CH1 cells.Methods The expressions of miR-519e-5p and ANXA5 in human chordoma tissues and adjacent tissues were detected by qRT-PCR.U-CH1 cells were cultured in vitro and divided into miR-519e-5p mimics group,miR-519e-5p mimics negative control+empty vector group,miR-519e-5p mimics+ANXA5 overexpression group,and control group.The proliferation of U-CH1 cells in each group was detected by plate colony formation assay and MTT assay;flow cytometry was used to detect the apoptosis of U-CH1 cells in each group;the migration and invasion of U-CH1 cells in each group were detected by scratch would healing assay and Transwell assay respectively;the expression of ANXA5 protein,apoptosis-related proteins(Bcl-2,Bax)and migration epithelial-mesenchymal transition-related proteins(Claudin-1,Vimentin,E-cadherin)of U-CH1 cells in each group were detected by Western blot;the expression of miR-519e-5p and ANXA5 mRNA of U-CH1 cells in each group was detected by qRT-PCR;and the interaction between miR-519e-5p and ANXA5 in U-CH1 cells was examined by dual-luciferase reporter gene assay.Results Compared with the adjacent tissues,the expression of miR-519e-5p in chordoma tissues decreased(P<0.05),and the expression of ANXA5 mRNA increased(P<0.05).Compared with the control group,the cell proliferation rate,colony formation rate,migration rate,invasion number,expressions of ANXA5 mRNA and ANXA5,Bcl-2,Claudin-1,and Vimentin proteins in the miR-519e-5p mimics group decreased(P<0.05),the expression of miR-519e-5p mRNA,the apoptosis rate,and the expression of Bax and E-cadherin proteins increased(P<0.05).Compared with the miR-519e-5p mimics group,the colony formation rate,cell proliferation rate,invasion number,migration rate,expressions of ANXA5 mRNA and ANXA5,Bcl-2,Vimentin,and Claudin-1 proteins in the miR-519e-5p mimics+ANXA5 overexpression group increased(P<0.05),and the apoptosis rate,the expression of Bax and E-cadher

关 键 词：miR-519e-5p ANXA5 脊索瘤 U-CH1细胞 生物学行为 

分 类 号：R738.1[医药卫生—肿瘤]