Sun Xin, Liu Bin, Zhao Xu-Dong, Wang Li-Yin, Ji Wen-Yue
Department of Otorhinolaryngology, Shengjing Hospital, China Medical University, Heping, Shenyang, Liaoning 110004, P.R. China.
Oncol Rep. 2015 Mar;33(3):1221-6. doi: 10.3892/or.2015.3714. Epub 2015 Jan 13.
Laryngeal cancer is one of the most commonly occurring malignant cancers of the head and neck region. In the present study, we investigated the roles of miR-221 in laryngeal squamous cell carcinoma cell line, Hep-2. We examined the function and mechanism of miR-221 in Hep-2 cells using techniques of cell biology and molecular pathology, such as western blotting, quantitative PCR, immunohistochemical staining and flow cytometry. Using a luciferase assay, the apoptotic protease activating factor-1 (Apaf-1) mRNA 3'-UTR was shown to have complementary binding sites using bioinformatics prediction software including TargetScan, PicTar and miRanda. In conclusion, our results showed that miR-221 inhibition caused elevated expression levels of the Apaf-1 apoptotic pathway proteins caspase-3, -8 and -9. miR-221 may therefore be used as a novel therapeutic target for laryngeal cancer.
喉癌是头颈部最常见的恶性肿瘤之一。在本研究中，我们调查了miR-221在喉鳞状细胞癌细胞系Hep-2中的作用。我们使用细胞生物学和分子病理学技术，如蛋白质免疫印迹法、定量PCR、免疫组织化学染色和流式细胞术，研究了miR-221在Hep-2细胞中的功能和机制。使用荧光素酶报告基因检测法，通过包括TargetScan、PicTar和miRanda在内的生物信息学预测软件显示凋亡蛋白酶激活因子-1（Apaf-1）mRNA的3'-UTR具有互补结合位点。总之，我们的结果表明，抑制miR-221会导致Apaf-1凋亡途径蛋白caspase-3、-8和-9的表达水平升高。因此，miR-221可能用作喉癌的新型治疗靶点。
