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

Let-7a inhibits migration and invasion of human oral squamous cell carcinoma cells by targeting matrix metalloproteinase-11

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。

Objective: To investigate the expression of let-7 a in oral squamous cell carcinoma and explore the effect of let-7 a 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-7 a and MMP11 by RT-qPCR and Western blot. Cell migration and invasion of OSCC-15 and HSC3 cells after over-expression of let-7 a or knockdown of MMP11 were detected by Transwell. The regulating relationship between let-7 a and MMP11 was verified by TargetScan online prediction, dual luciferase reporter system and Western blot. Cell migration and invasion after over-expression of let-7 a or co-expression of let-7 a and MMP11 were detected by Transwell. Results: The expression of let-7 a 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-7 a was negatively correlated with MMP11(r^2=0.6359, P<0.0001). The migration and invasion of OSCC-15 and HSC3 cells were inhibited after over-expression of let-7 a or knockdown of MMP11. TargetScan online prediction and dual luciferase assay indicates that MMP11 is a potential target gene for let-7 a and let-7 a negatively regulated the expression of MMP11. MMP11 could rescue the inhibitory effect of let-7 a on migration and invasion of OSCC-15 and HSC3 cells. Conclusion: Let-7 a is downregulated while MMP11 is up-regulated in oral squamous cell carcinoma, and let-7 a can inhibit the migration and invasion of oral squamous cell carcinoma cell lines OSCC-15 and HSC3 by targeting MMP11.