华西口腔医学杂志 ›› 2020, Vol. 38 ›› Issue (2): 185-192.doi: 10.7518/hxkq.2020.02.013

• 口腔肿瘤学专栏 • 上一篇    下一篇

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

郑晶, 张雨温, 李天客, 包阳, 张素欣()   

  1. 河北医科大学第四医院口腔科,石家庄 050011
  • 收稿日期:2019-07-17 修回日期:2020-01-12 出版日期:2020-04-01 发布日期:2020-04-15
  • 通讯作者: 张素欣 E-mail:suxin316@163.com
  • 作者简介:郑晶,主治医师,硕士,E-mail:wangguo888@sina.com
  • 基金资助:
    河北省卫生健康委员会重点科技研究计划项目(201906-95)

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()   

  1. Dept. of Stomatology, The Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China
  • Received:2019-07-17 Revised:2020-01-12 Online:2020-04-01 Published:2020-04-15
  • Contact: Suxin Zhang E-mail:suxin316@163.com
  • Supported by:
    Hebei Health and Fitness Commission Key Science and Technology Research Plan Project(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

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.

Key words: miR-204-5p, bromodomain-containing protein 4, proliferation, migration and invasion

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