Department of Oral and Maxillofacial Surgery, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8571, Japan.
Department of Oral and Maxillofacial Surgery, Faculty of Medicine, University of Tsukuba, Tsukuba, Ibaraki 305-8571, Japan.
Int J Oncol. 2018 Mar;52(3):841-850. doi: 10.3892/ijo.2018.4260. Epub 2018 Jan 31.
MicroRNAs (miRNAs or miRs) play important roles in carcinogenesis. The miRNA, miR-205-5p, has been reported to suppress the growth of various types of tumor; however, its functional contribution to oral squamous cell carcinoma (OSCC) is not yet clear. Thus, this study was conducted to determine the miRNA expression signatures in OSCC and to investigate the functional role of miR‑205‑5p in OSCC cells. We measured miR‑205‑5p expression by RT-qPCR, and examined the function of miR‑205‑5p by transfecting a miR‑205‑5p mimic or inhibitor into OSCC cells and measuring cell proliferation, migration and invasiveness. Genes targeted by miR‑205‑5p were identified using the TargetScan database and verified by western blot analysis, luciferase reporter assay and ELISA. We found that miR‑205‑5p was significantly downregulated in OSCC cell lines and tissue specimens. Following transfection of miR‑205‑5p mimic or inhibitor into the cancer cell lines, miR‑205‑5p overexpression significantly suppressed cancer cell migration and invasion. We further demonstrated that miR‑205‑5p directly targeted and regulated the tissue inhibitor of metalloproteinases‑2 (TIMP‑2) gene. The silencing of TIMP‑2 suppressed cancer cell invasion and the activation of pro‑matrix metalloproteinase‑2 (pro‑MMP‑2). These results suggest that TIMP‑2 promotes tumor progression, and that miR‑205‑5p directly regulates TIMP‑2, thereby suppressing pro‑MMP‑2 activation and inhibiting OSCC cell invasiveness. Our data describing the pathways regulated by miR‑205‑5p provide new insight into the mechanisms responsible for OSCC development and metastasis.
微小 RNA(miRNA 或 miR)在致癌作用中发挥重要作用。有报道称 miRNA,miR-205-5p,可抑制多种类型肿瘤的生长;然而,其在口腔鳞状细胞癌(OSCC)中的功能贡献尚不清楚。因此,本研究旨在确定 OSCC 中的 miRNA 表达谱,并研究 miR-205-5p 在 OSCC 细胞中的功能作用。我们通过 RT-qPCR 测量 miR-205-5p 的表达,并通过转染 miR-205-5p 模拟物或抑制剂到 OSCC 细胞中,测量细胞增殖、迁移和侵袭来检查 miR-205-5p 的功能。使用 TargetScan 数据库鉴定 miR-205-5p 靶向的基因,并通过 Western blot 分析、荧光素酶报告基因测定和 ELISA 进行验证。我们发现 miR-205-5p 在 OSCC 细胞系和组织标本中显著下调。在将 miR-205-5p 模拟物或抑制剂转染入癌细胞系后,miR-205-5p 的过表达显著抑制了癌细胞的迁移和侵袭。我们进一步表明,miR-205-5p 直接靶向和调节组织金属蛋白酶抑制剂-2(TIMP-2)基因。TIMP-2 的沉默抑制了癌细胞的侵袭和前基质金属蛋白酶-2(pro-MMP-2)的激活。这些结果表明 TIMP-2 促进肿瘤进展,miR-205-5p 直接调节 TIMP-2,从而抑制 pro-MMP-2 激活并抑制 OSCC 细胞侵袭。我们描述 miR-205-5p 调节的途径的数据为 OSCC 发展和转移的机制提供了新的见解。