中国药业2024,Vol.33Issue(18) :26-33.DOI:10.3969/j.issn.1006-4931.2024.18.007

miRNA-4465靶向Wnt5A调控对胶质瘤细胞增殖、迁移和侵袭的影响

Effects of Regulation of miRNA-4465 Targeting Wnt5A on the Proliferation,Migration and Invasion of Glioma Cells

陈硕硕 孙玉玺 余德 束汉生
中国药业2024,Vol.33Issue(18) :26-33.DOI:10.3969/j.issn.1006-4931.2024.18.007

miRNA-4465靶向Wnt5A调控对胶质瘤细胞增殖、迁移和侵袭的影响

Effects of Regulation of miRNA-4465 Targeting Wnt5A on the Proliferation,Migration and Invasion of Glioma Cells

陈硕硕 1孙玉玺 1余德 1束汉生1
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作者信息

  • 1. 蚌埠医学院第二附属医院,安徽 蚌埠 233000
  • 折叠

摘要

目的 探讨miRNA-4465(miR-4465)对胶质瘤细胞增殖、侵袭及迁移的影响及其作用机制.方法 采用miR-4465 模拟物(mimic)处理U118 和U87 细胞,检测细胞生物活性.双荧光素酶报告基因实验评价miR-4465 和Wnt5A的结合情况.采用siRNA和miR-4465 抑制剂(inhibitor)分别抑制Wnt5A和miR-4465 的表达,采用CCK-8 法、细胞克隆实验、伤口愈合实验和Transwell实验,检测细胞的增殖、迁移、侵袭能力.复制移植瘤裸鼠模型,评价miR-4465 过表达和抑制对其肿瘤生长的影响.结果 miR-4465 mimic显著抑制了U118 和U87 细胞增殖能力.双荧光素酶报告基因实验结果显示miR-4465 与Wnt5A存在靶向调节作用.沉默Wnt5A可降低细胞的增殖、迁移、侵袭能力,miR-4465 被抑制后,细胞的增殖、迁移、侵袭能力均显著增强(P<0.05).此外,沉默Wnt5A可逆转miR-4465 抑制导致的细胞增殖、迁移、侵袭能力增强.体内实验结果显示,miR-4465 过表达抑制肿瘤生长和ki67 的表达,而miR-4465 被抑制后,肿瘤生长和ki67 的表达显著增强(P<0.05).结论 miR-4465 在胶质瘤中呈低表达,其过表达可抑制胶质瘤细胞的增殖、迁移、侵袭能力,其下游作用机制与Wnt5A通路有关.

Abstract

Objective To investigate the effects of miRNA-4465(miR-4465)on the proliferation,invasion,and migration of glioma cells and relevant mechanism.Methods The U118 and U87 cells were treated by the miR-4465 mimic to detect the cell biological activity.The binding of miR-4465 and Wnt5A was evaluated by the dual-luciferase reporter gene test.The expression of Wnt5A and miR-4465 was inhibited by the siRNA and miR-4465 inhibitors,respectively.The proliferation,migration and invasion abilities of cells were detected by the CCK-8 method,clone formation assay,wound healing assay and Transwell assay.The effects of overexpression and inhibition of miR-4465 on tumor growth were evaluated by constructing transplanted-tumor nude mice model.Results The miR-4465 mimic significantly inhibited the proliferation ability of U118 and U87 cells.The dual-luciferase reporter gene test showed a targeted regulatory effect between miR-4465 and Wnt5A.Wnt5A silence could decrease the proliferation,migration and invasion abilities of cells,the above abilities of cells significantly increased after miR-4465 inhibition(P<0.05).In addition,Wnt5A silence could reversed the increased cell proliferation,migration and invasion abilities induced by miR-4465 inhibition.The in vivo tests showed that overexpression of miR-4465 inhibited the tumor growth and ki67 expression,while the inhibition of miR-4465 promoted tumor growth and ki67 expression(P<0.05).Conclusion The expression of miR-4465 is low in gliomas,and its overexpression can inhibit the proliferation,migration and invasion abilities of glioma cells.Its downstream mechanism is related to the Wnt5A pathway.

关键词

胶质瘤/miR-4465/双荧光素酶报告基因/细胞/增殖/迁移/侵袭/Wnt5A

Key words

glioma/miR-4465/dual-luciferase reporter gene/cell/proliferation/migration/invasion/Wnt5A

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基金项目

安徽省高校自然科学研究项目(KJ2021A0786)

出版年

2024
中国药业
重庆市食品药品监督管理局

中国药业

CSTPCD
影响因子:1.369
ISSN:1006-4931
参考文献量5
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