Effect of miR-519e-5p targeting ANXA5 on the biological behavior of chordoma U-CH1 cells
- JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY Vol. 33, Issue 10, Pages: 892-897(2024)
- 作者机构：
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
- 作者简介：
- 基金信息：
DOI：10.11659/jjssx.05E023060
CLC： R738.1Published：25 October 2024，
Received：12 May 2023，
- 稿件说明：
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李轶,黄小平,王玮.miR-519e-5p靶向ANXA5对脊索瘤U-CH1细胞生物学行为的影响[J].局解手术学杂志,2024,33(10):892-897.
LI Yi,HUANG Xiao-ping,WANG Wei.Effect of miR-519e-5p targeting ANXA5 on the biological behavior of chordoma U-CH1 cells[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2024,33(10):892-897.
李轶,黄小平,王玮.miR-519e-5p靶向ANXA5对脊索瘤U-CH1细胞生物学行为的影响[J].局解手术学杂志,2024,33(10):892-897. DOI： 10.11659/jjssx.05E023060.
LI Yi,HUANG Xiao-ping,WANG Wei.Effect of miR-519e-5p targeting ANXA5 on the biological behavior of chordoma U-CH1 cells[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2024,33(10):892-897. DOI： 10.11659/jjssx.05E023060.
摘要
目的
2
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
2
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
2
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
2
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。
Abstract
Objective
2
To investigate the effect of miR-519e-5p targeting Annexin A5 （ANXA5） on the biological behavior of chordoma U-CH1 cells.
Methods
2
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
2
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-cadherin proteins decreased （
P
<
0.05）. MiR-519e-5p can target and down-regulate the expression of ANXA5 in U-CH1 cells.
Conclusion
2
MiR-519e-5p reduces the migration， proliferation and invasion viability of chordoma U-CH1 cells， and promotes apoptosis by targeting the inhibition of ANXA5 expression.
关键词
miR-519e-5pANXA5脊索瘤U-CH1细胞生物学行为
Keywords
miR-519e-5pANXA5chordomaU-CH1 cellsbiological behavior
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