生物化学与生物物理进展2024,Vol.51Issue(5) :1203-1214.DOI:10.16476/j.pibb.2023.0391

miR-375靶向MMP13抑制骨肉瘤细胞的迁移和侵袭

miR-375 Attenuates The Migration and Invasion of Osteosarcoma Cells by Targeting MMP13

刘中 何磊 肖剑 朱青梅 肖君 杨勇明 罗勇健 莫中成 张轶群 李明
生物化学与生物物理进展2024,Vol.51Issue(5) :1203-1214.DOI:10.16476/j.pibb.2023.0391

miR-375靶向MMP13抑制骨肉瘤细胞的迁移和侵袭

miR-375 Attenuates The Migration and Invasion of Osteosarcoma Cells by Targeting MMP13

刘中 1何磊 1肖剑 1朱青梅 1肖君 1杨勇明 1罗勇健 1莫中成 2张轶群 1李明3
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作者信息

  • 1. 邵阳学院附属第一医院骨科,邵阳 422000
  • 2. 邵阳学院附属第一医院骨科,邵阳 422000;桂林医学院基础医学院,桂林 541199
  • 3. 湖南医药学院基础医学院,怀化 418000
  • 折叠

摘要

目的 为了探究miR-375是否通过影响基质金属蛋白酶13(MMP13)的表达来调控骨肉瘤(osteosarcoma,OS)恶性特征.方法 用Lipofectamine 3000试剂盒将质粒、miRNA转染至骨肉瘤细胞和HEK293细胞中.实时定量聚合酶链反应(real-time quantitative PCR,RT-qPCR)检测 OS患者和OS细胞中 miR-375 和 MMP13 的表达.蛋白质印迹法(Western blot)分析OS患者和OS细胞中MMP13蛋白的表达.双荧光素酶法分析miR-375与MMP13的靶向关系.伤口愈合和transwell实验分别分析OS细胞的迁移和侵袭.结果 OS组织中miR-375的表达低于正常组织.MMP13在OS组织中表达上调.在OS患者中,MMP13的表达与miR-375呈负相关.与转染miRNA对照的OS细胞相比,转染miR-375模拟物OS细胞的迁移和侵袭明显被抑制.MMP13能部分逆转miR-375对OS细胞迁移和侵袭的抑制作用.结论 在OS细胞中,过表达miR-375通过调控MMP13的表达抑制细胞的迁移和侵袭.

Abstract

Objective To explore whether miR-375 regulates the malignant characteristics of osteosarcoma(OS)by influencing the expression of MMP13.Methods Plasmid DNAs and miRNAs were transfected into OS cells and HEK293 cells using Lipofectamine 3000 reagent.Real-time quantitative polymerase chain reaction was performed to measure the expression of miR-375 and MMP13 in OS patients and OS cells.Western blot was performed to analyze the MMP13 protein in the patients with OS and OS cells.The targeting relationship between miR-375 and MMP13 was analyzed by luciferase assay.Migration and invasion were analysed by heal wound and transwell assays,respectively.Results miR-375 expression in OS tissues was lower than that in normal tissues.The expression of MMP13 was upregulated in OS tissues.MMP13 expression was negatively correlated with miR-375 expression in patients with OS.Migration and invasion were significantly inhibited in OS cells with the miR-375 mimic compared with OS cells with the miRNA control.MMP13 partially reversed the inhibition of migration and invasion induced by miR-375 in the OS cells.Conclusion miR-375 attenuates migration and invasion by downregulating the expression of MMP13 in OS cells.

关键词

骨肉瘤/miR-375/基质金属蛋白酶13/细胞迁移/细胞侵袭

Key words

osteosarcoma/miR-375/MMP13/migration/invasion

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基金项目

湖南省自然科学基金(2021JJ30482)

湖南省自然科学基金(2022JJ50192)

邵阳市科技计划(2020GZ48)

邵阳学院附属第一医院横向合作项目(2020HX03)

出版年

2024
生物化学与生物物理进展
中国科学院生物物理研究所,中国生物物理学会

生物化学与生物物理进展

CSTPCD北大核心
影响因子:0.476
ISSN:1000-3282
参考文献量35
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