miR-204-5p靶向溴结构域蛋白4对舌鳞状细胞癌细胞SCC25增殖和迁移侵袭的影响研究

Effect of miR-204-5p on the proliferation,migration,and invasion on tongue squamous cell carcinoma SCC25 cells by targeting bromodomain-containing protein 4

目的探讨miR-204-5p对溴结构域蛋白(BRD)4的靶向作用机制以及对舌鳞状细胞癌SCC25细胞增殖、迁移和侵袭的影响。方法实时荧光定量聚合酶链式反应(RT-qPCR)检测舌鳞状细胞癌组织以及不同细胞株系间的miR-204-5p和BRD4 mRNA的表达水平;miR-204-5p转染SCC25细胞后,细胞计数试剂盒(CCK)8法检测其对细胞增殖的影响,Transwell小室法检测miR-204-5p对SCC25迁移和侵袭的影响;使用TargetScan和荧光素酶实验分析并验证miR-204-5p与BRD4的靶向调控关系;RT-qPCR和Western blot检测miR-204-5p模拟物和抑制剂对BRD4表达水平的影响;Transwell小室法和CCK8法检测miR-204-5p调控BRD4对SCC25增殖和迁移侵袭的影响。结果miR-204-5p在舌鳞状细胞癌组织和SCC25细胞中表达下调,BRD4在舌鳞状细胞癌组织和SCC25细胞中表达上调;提高miR-204-5p表达量抑制SCC25细胞的增殖、迁移和侵袭;TargetScan和荧光素酶实验证实miR-204-5p与BRD4具有靶向负调控关系;miR-204-5p模拟物抑制BRD4表达,miR-204-5p抑制剂促进BRD4表达上调;当在SCC25细胞中同时过表达miR-204-5p和BRD4时,BRD4能够回补miR-204-5p对SCC25细胞的抑制作用。结论miR-204-5p能够通过靶向负调控BRD4抑制舌鳞状细胞癌SCC25细胞的增殖、迁移和侵袭。

Objective This study aimed to explore the target relationship between miR-204-5p and bromodomain-containing protein(BRD)4,as well as their effects on cell proliferation,migration,and invasion in tongue squamous cell carcinoma SCC25.Methods Real-time quantitative polymerase chain reaction(RT-qPCR)was performed to detect miR-204-5p and BRD4 expression levels in tongue squamous cell carcinoma and different cell lines.TargetScan and dual luciferase reporter assay were used to confirm the target relationship between miR-204-5p and BRD4.The effects of miR-204-5p on SCC25 cell proliferation were examined by cell counting kit(CCK)8 assay,whereas those on SCC25 cell migration and invasion were determined by Transwell assay.RT-qPCR and Western blot were used to detect the effects of miR-204-5p mimics and inhibitors on BRD4 expression.Transwell and CCK8 assays were used to detect the effects of miR-204-5p on proliferation,migration,and invasion through BRD4 regulation.Results miR-204-5p was significantly downregulated in the tissues and cells of squamous cell carcinoma,and BRD4 showed the opposite result.The increase in miR-204-5p expression can inhibit the proliferation,migration,and invasion of SCC25 cells.TargetScan and luciferase test confirmed that miR-204-5p and BRD4 had a negative regulatory relationship with BRD4,respectively.Moreover,miR-204-5p mimics can inhibit BRD4 expression,and miR-204-5p inhibitors can promote BRD4 expression upregulation.When miR-204-5p and BRD4 were overexpressed in SCC25 cells,BRD4 can make up for the inhibitory effect of miR-204-5p on SCC25 cells.Conclusion miR-204-5p could inhibit proliferation,migration and invasion in tongue squamous cell carcinoma SCC25 cells by targeting BRD4 gene.