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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
引用本文:郑晶,张雨温,李天客,包阳,张素欣.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[J].华西口腔医学杂志,2020,38(2):185-192.
作者姓名:郑晶  张雨温  李天客  包阳  张素欣
作者单位:河北医科大学第四医院口腔科,石家庄 050011;河北医科大学第四医院口腔科,石家庄 050011;河北医科大学第四医院口腔科,石家庄 050011;河北医科大学第四医院口腔科,石家庄 050011;河北医科大学第四医院口腔科,石家庄 050011
基金项目:河北省卫生健康委员会重点科技研究计划项目(201906-95)
摘    要: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。

关 键 词:miR-204-5p  溴结构域蛋白4  增殖  迁移侵袭
收稿时间:2019-07-17
修稿时间:2020-01-12

Effect of miR-204-5p on the proliferation,migration, and invasion on tongue squamous cell carcinoma SCC25 cells by targeting bromodomain-containing protein 4
Zheng Jing,Zhang Yuwen,Li Tianke,Bao Yang,Zhang Suxin.Effect of miR-204-5p on the proliferation,migration, and invasion on tongue squamous cell carcinoma SCC25 cells by targeting bromodomain-containing protein 4[J].West China Journal of Stomatology,2020,38(2):185-192.
Authors:Zheng Jing  Zhang Yuwen  Li Tianke  Bao Yang  Zhang Suxin
Institution:Dept. of Stomatology, The Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China
Abstract: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.
Keywords:miR-204-5p  bromodomain-containing protein 4  proliferation  migration and invasion  
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