Department of Thoracic and Cardiovascular Surgery, The First Affiliated Hospital of Soochow University, Medical College of Soochow University Suzhou, China ; Soochow University Laboratory of Cancer Molecular Genetics, Medical College of Soochow University Suzhou, China ; Suzhou Key Laboratory for Cancer Molecular Genetics Suzhou, China.
Department of Thoracic and Cardiovascular Surgery, The First Affiliated Hospital of Soochow University, Medical College of Soochow University Suzhou, China.
Thorac Cancer. 2015 Jul;6(4):524-33. doi: 10.1111/1759-7714.12242. Epub 2015 Feb 27.
Accumulated evidence has revealed that the dysregulation of micro ribonucleic acids (miRNAs) may contribute to esophageal squamous cell carcinoma (ESCC). MiR-93, which is a member of the miRNA cluster miR-106b∼25, has been widely studied for its tumor promoting effect on different types of cancers. However, our knowledge of miR-93 function in ESCC remains unclear.
The expression levels of miR-93 in ESCC and the adjacent non-tumor tissues were measured by real-time polymerase chain reaction. Cell counting kit-8, flow cytometry, and 5-ethynyl-2'-deoxyuridine incorporation and transwell migration assays were employed to explore the effects of miR-93 on proliferation and migration capabilities in EC109 cells. To determine the possible target gene of miR-93, cell transfection, Western blot analysis and luciferase reporter gene assays were performed.
A significant upregulation of miR-93 expression in ESCC tissues was determined, combined with a downregulation of the predicted target gene, disabled 2 (DAB2). The introduction of miR-93 significantly promotes cell proliferation, cell cycle progression, and the metastatic capability of EC109 cells. By cell transfection and luciferase reporter assay, DAB2 was confirmed as a direct target of miR-93. In addition, the knockdown of DAB2 by small interfering RNA displayed a consentaneous phenocopy with miR-93 overexpression in EC109 cells.
Our results indicate that miR-93 acts as a tumor promoter in ESCC, and its promotion effects on ESCC cell proliferation and migration depend largely upon DAB2 suppression.
已有大量证据表明微小核糖核酸(miRNAs)的失调可能导致食管鳞状细胞癌(ESCC)。miR-93 是 miRNA 簇 miR-106b∼25 的成员,其在不同类型癌症中的肿瘤促进作用已得到广泛研究。然而,我们对 miR-93 在 ESCC 中的功能知之甚少。
通过实时聚合酶链反应测量 ESCC 和相邻非肿瘤组织中 miR-93 的表达水平。使用细胞计数试剂盒-8、流式细胞术、5-乙炔基-2'-脱氧尿苷掺入和 Transwell 迁移实验来探索 miR-93 对 EC109 细胞增殖和迁移能力的影响。为了确定 miR-93 的可能靶基因,进行了细胞转染、Western blot 分析和荧光素酶报告基因检测。
确定 ESCC 组织中 miR-93 的表达显著上调,同时预测的靶基因Disabled 2(DAB2)下调。miR-93 的引入显著促进了 EC109 细胞的增殖、细胞周期进程和转移能力。通过细胞转染和荧光素酶报告基因检测,确认 DAB2 是 miR-93 的直接靶基因。此外,通过小干扰 RNA 敲低 DAB2 在 EC109 细胞中显示出与 miR-93 过表达一致的表型。
我们的结果表明,miR-93 在 ESCC 中作为肿瘤促进因子发挥作用,其对 ESCC 细胞增殖和迁移的促进作用在很大程度上依赖于 DAB2 的抑制。