Let-7a通过靶向基质金属蛋白酶-11抑制人口腔鳞状细胞癌细胞迁移和侵袭
| 作者: |
1魏薇,
1唐祎,
1代婧,
1陈胡杰
1 荆州市中心医院口腔科，湖北 荆州 434000 |
| 通讯: |
唐祎
Email: 20984688@qq.com |
| DOI: | 10.3978/j.issn.2095-6959.2018.11.003 |
| 基金: | 湖北省卫生计划项目(WJ2017M242)。 |
摘要
目的：探讨let-7a在口腔鳞状细胞癌组织中的表达，并研究let-7a靶向MMP11抑制口腔鳞状细胞癌细胞迁移和侵袭的作用机制。方法：收集口腔鳞状细胞癌患者的肿瘤组织及正常牙龈黏膜组织，采用RT-qPCR和Western印迹法检测组织样本及口腔鳞状细胞癌细胞系中let-7a和MMP11相对表达水平；以OSCC-15，HSC3细胞为研究对象，Transwell法分别检测转染let-7a mimics、敲低MMP11对细胞迁移和侵袭的影响；采用TargetScan在线预测、双荧光素酶报告基因实验和Western印迹法验证let-7a与MMP11的靶向关系；过表达let-7a或共表达let-7a和MMP11，Transwell法检测细胞迁移和侵袭。结果：对比正常牙龈黏膜组织，口腔鳞状细胞癌组织中let-7a表达明显下调(P<0.05)，MMP11表达明显上调(P<0.05)，let-7a与MMP11表达呈负相关(r2=0.6359，P<0.0001)；在OSCC-15和HSC3细胞中，过表达let-7a或敲减MMP11能明显抑制细胞迁移和侵袭(P<0.05)。经TargetScan在线预测和双荧光素酶报告基因实验验证，MMP11是let-7a的潜在靶基因，let-7a调控MMP11表达；回补MMP11能够逆转let-7a对OSCC-15、HSC3细胞迁移和侵袭的抑制作用。结论：在口腔鳞状细胞癌组织中，let-7a表达下调，MMP11表达上调，let-7a能通过靶向MMP11抑制口腔鳞状细胞癌细胞株OSCC-15，HSC3细胞迁移和侵袭。
关键词：
let-7a；口腔鳞状细胞癌；基质金属蛋白酶-11；迁移；侵袭
Let-7a inhibits migration and invasion of human oral squamous cell carcinoma cells by targeting matrix metalloproteinase-11
CorrespondingAuthor: TANG Yi Email: 20984688@qq.com
DOI: 10.3978/j.issn.2095-6959.2018.11.003
Foundation: This work was supported by Health Project in Hubei Province, China (WJ2017M242).
Abstract
Objective: To investigate the expression of let-7a in oral squamous cell carcinoma and explore the effect of let-7a on migration and invasion of oral squamous cell carcinoma by targeting MMP11. Methods: Tumor tissues and normal gingival mucosa tissues of patients with oral squamous cell carcinoma were collected to detect the expression of let-7a and MMP11 by RT-qPCR and Western blot. Cell migration and invasion of OSCC-15 and HSC3 cells after over-expression of let-7a or knockdown of MMP11 were detected by Transwell. The regulating relationship between let-7a and MMP11 was verified by TargetScan online prediction, dual luciferase reporter system and Western blot. Cell migration and invasion after over-expression of let-7a or co-expression of let-7a and MMP11 were detected by Transwell. Results: The expression of let-7a in oral squamous cell carcinoma tissues was significantly lower than that in normal gingival mucosa tissues (P<0.05), while MMP11 had the opposite trend in the tissues (P<0.05). The expression of let-7a was negatively correlated with MMP11 (r2=0.6359, P<0.0001). The migration and invasion of OSCC-15 and HSC3 cells were inhibited after over-expression of let-7a or knockdown of MMP11. TargetScan online prediction and dual luciferase assay indicates that MMP11 is a potential target gene for let-7a and let-7a negatively regulated the expression of MMP11. MMP11 could rescue the inhibitory effect of let-7a on migration and invasion of OSCC-15 and HSC3 cells. Conclusion: Let-7a is down-regulated while MMP11 is up-regulated in oral squamous cell carcinoma, and let-7a can inhibit the migration and invasion of oral squamous cell carcinoma cell lines OSCC-15 and HSC3 by targeting MMP11.
Keywords：
let-7a; oral squamous cell carcinoma; matrix metalloproteinase-11; migration; invasion