摘要： 目的：探讨miR-149通过靶向调节基因对口腔鳞癌细胞生长和侵袭的影响。方法：以实验室过微阵列芯片技术筛选出的miR-149作为本实验研究对象，通过靶基因预测软件，选定基质金属蛋白酶-9（MMP-9）为其靶基因，利用实时定量PCR（RT-PCR）检测口腔鳞癌和癌旁组织中miR-149和MMP-9 mRNA的表达水平。瞬时转染miR-149 mimics、miR-149 mimics control、MMP-9siRNA和MMP-9 siRNA阴性对照，通过PT-PCR及Western blotting 检测口腔鳞癌KB细胞系中miR-149与MMP-9的关系。运用平板克隆形成和Transwell侵袭实验检测MMP-9对口腔鳞癌KB细胞生长和侵袭的影响。结果：在口腔鳞癌中miR-149的表达明显低于癌旁组织（P<0.01），MMP-9 mRNA的情况则相反（P<0.01）。瞬时转染后应用RT-PCR及Western blotting 检测提示，miR-149与MMP-9表达成反比，差异具有统计学意义（P＜0.05），说明二者之间有靶向调节关系。与对照组细胞相比，转染MMP-9 siRNA的口腔鳞癌KB细胞克隆数目减少（P<0.05），发生侵袭的细胞减少（P<0.05）。结论： miR-149通过靶向调节MMP-9的表达，抑制口腔鳞癌细胞的生长和侵袭，发挥着抑癌基因的作用，可望作为口腔鳞癌分子治疗的有效靶点。

关键词: 口腔鳞癌；  , 靶向调节；  , microRNA-149；  , 基质金属蛋白酶-9

Abstract: Objective: To explore the targeting regulatory impact of miR-149 target genes  on oral squamous cell carcinoma (OSCC) cell growth and invasion. Methods: Laboratory used microarray screening of miR-149 as an object of study in this experiment. Matrix metalloproteinase -9 (MMP-9) selected by the target gene prediction software were used as target genes.  RT-PCR was performed to detect miR-149 and MMP-9 mRNA expression levels in OSCC and tumor adjacent tissues. Transiently transfected with miR-149 mimics, miR-149 mimics control, MMP-9 siRNA and MMP-9 siRNA negative control, PT-PCR and Western blotting were performed to examine the correlation of miR-149 and MMP-9 expressions in OSCC KB cell line. MMP-9 in OSCC KB cell growth and invasion assays were performed with the use of colony formation assay and Transwell chambers. Results: In OSCC, miR-149 expression was significantly lower than adjacent tissues (P<0.01). But MMP-9 mRNA was significantly higher than adjacent tissue (P<0.01).  The expression of miR- 149 and MMP-9 revealed a negative correlation examined by RT-PCR and Western blotting after transient transfection, the difference was significant (P<0.05). As compared with control cells, the cell clone numbers of OSCC KB cells transfected by  MMP-9 siRNA decreased (P<0.05), and the number of invasive cells in this group decreased also (P<0.05). Conclusion: miR-149 inhibit the growth and invasion of OSCC by target regulation of MMP-9 expression.

Key words: oral squamous cell carcinoma, targeted adjustment, microRNA-149, matrix metalloptoteinases-9

中图分类号: 

• R782.2