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,Migrationand Invasion of Glioma Cells

作　　者：陈硕硕 孙玉玺 余德 束汉生[1] CHEN Shuoshuo;SUN Yuxi;YU De;SHU Hansheng(The Second Affiliated Hospital of Bengbu Medical College,Bengbu,Anhui,China 233000)

机构地区：[1]蚌埠医学院第二附属医院,安徽蚌埠233000

出　　处：《中国药业》2024年第18期26-33,共8页China Pharmaceuticals

基　　金：安徽省高校自然科学研究项目[KJ2021A0786]。

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。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 

分 类 号：R965[医药卫生—药理学] R739.41[医药卫生—药学]