微小RNA-483-5p调控TIMP2表达对膀胱癌细胞增殖和侵袭的影响

Targeted-regulation of TIMP2 expression by microRNA-483-5p and its effect on the proliferation and invasion of bladder cancer cells

目的探讨微小RNA-483-5p(miR-483-5p)对膀胱癌(BC)细胞增殖、迁移和侵袭的影响,并初步分析其可能的分子机制。方法通过TCGA数据库分析miR-483-5p在BC组织中的表达及与预后的关系,荧光实时定量PCR检测BC细胞T24中miR-483-5p和金属蛋白酶组织抑制因子(TIMP)-2的表达水平。将miR-483-5p模拟物(mimic)和阻碍物(inhibitor)转染T24细胞,应用CCK-8和Transwell法评估miR-483-5p对T24细胞增殖和迁移侵袭的影响。双荧光素酶报告基因实验分析miR-483-5p和TIMP-2间的靶向关系,挽救实验验证miR-483-5p的生物学功能是否通过TIMP-2发挥作用。结果与癌旁组织比较,BC组织中高表达miR-483-5p(P<0.05),高表达miR-483-5p者的总生存率低于低表达者(P<0.05)。miR-483-5p mimic可促进T24细胞的增殖、迁移和侵袭能力(P<0.05),miR-483-5p inhibitor则抑制T24细胞的增殖、迁移和侵袭能力(P<0.05)。TIMP2为miR-483-5p的下游靶点,干扰TIMP2可逆转miR-483-5p下调对T24细胞增殖、迁移和侵袭的抑制作用(P<0.05)。结论miR-483-5p在BC中高表达,与BC患者不良预后相关。miR-483-5p可下调TIMP2的表达来促进BC细胞的增殖、迁移和侵袭。

Objective To investigate the impact of microRNA-483-5p(miR-483-5p)on the proliferation,migration,and invasion of bladder cancer(BC)cells as well as its potential molecular mechanism.Methods The expression of miR-483-5p in BC tissues and its relationship with prognosis were analyzed by TCGA database.Fluorescence-based real-time quantitative PCR was used to analyze the expression levels of miR-483-5p and tissue inhibitor of matrix metalloproteinases(TIMP)-2 in BC T24 cells.Transfection of miR-483-5p mimic and inhibitor was performed in T24 cells.The impact of miR-483-5p on cellular proliferation was evaluated through the implementation of a CCK-8 assay.Migration and invasion capabilities of T24 cells were assessed utilizing Transwell assays.The targeted interaction between miR-483-5p and TIMP-2 was demonstrated using a dual-luciferase reporter gene assay.Rescue experiments was performed to verify whether the biological functions of miR-483-5p play a role through TIMP-2.Results The expression of miR-483-5p in BC tissues was significantly higher compared to adjacent tissues(P<0.05).Patients with high expression of miR-483-5p exhibited a lower overall survival rate than those with low expression(P<0.05).The miR-483-5p mimic significantly enhanced the proliferation,migration,and invasion capacities of T24 cells(P<0.05).The inhibition of miR-483-5p resulted in a significant decrease in proliferation,migration and invasion capacities of T24 cells(P<0.05).The downstream targeted gene of miR-483-5p,TIMP2,was found to be involved in regulating proliferation,migration and invasion capacities of T24 cells(P<0.05).Furthermore,the downregulation effects of miR-483-5p on these cellular processes can be reversed by using interfering TIMP2.Conclusion The expression of miR-483-5p is upregulated in BC and correlates with unfavorable prognosis in BC patients.Moreover,miR-483-5p exerts its oncogenic effects by suppressing the expression of TIMP2,thereby promoting proliferation,migration,and invasion of BC cells.