《Journal of Oral and Maxillofacial Surgery》 ›› 2015, Vol. 25 ›› Issue (5): 321-. doi: 10.3969/j.issn.1005-4979.2015.05.002

• Basic Scientific Study • Previous Articles     Next Articles

Expression of miR-149  in Oral Squamous Cell Carcinoma KB Cells and its Targeting Regulation of MMP-9

PEI Hao   

  1. Department of Stomatology, Nanyang Medical College First Hospital, Nanyang 473058, Henan Province, China
  • Online:2015-10-28 Published:2015-12-17
  • Supported by:

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miR-149 在口腔鳞癌细胞株KB中的表达及其对MMP-9的靶向调节作用

裴浩   

  1. 河南省南阳医学高等专科学校第一附属医院口腔科,河南   南阳   473058
  • 通讯作者: 裴浩,副主任医师 E-mail:peihao1227@163.com
  • 作者简介:裴浩(1969—),男,济南南阳人,学士,副主任医师
  • 基金资助:

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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

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

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