BACKGROUND MicroRNA-720 (miR-720), a nonclassical miRNA, is involved in the initiation and progression of several tumors. In our previous studies, miR-720 was shown to be significantly upregulated in cervical cancer tissues compared with normal cervical tissues. However, the precise biological functions of miR-720, and its molecular mechanisms of action, are still unknown. RESULTS Microarray expression profiles, luciferase reporter assays, and western blot assays were used to validate Rab35 as a target gene of miR-720 in HEK293T and HeLa cells. The regulation of Rab35 expression by miR-720 was assessed using qRT-PCR and western blot assays, and the effects of exogenous miR-720 and Rab35 on cell migration were evaluated in vitro using Transwell(®) assay, wound healing assay, and real-time analyses in HeLa cells. The influences of exogenous miR-720 on cell proliferation were evaluated in vitro by the MTT assay in HeLa cells. In addition, expression of E-cadherin and vimentin associated with epithelial-mesenchymal transition were also assessed using western blot analyses after transfection of miR-720 mimics and Rab35 expression vectors. The results showed that the small GTPase, Rab35, is a direct functional target of miR-720 in cervical cancer HeLa cells. By targeting Rab35, overexpression of miR-720 resulted in a decrease in E-cadherin expression and an increase in vimentin expression and finally led to promotion of HeLa cell migration. Furthermore, reintroduction of Rab35 3'-UTR(-) markedly reversed the induction of cell migration in miR-720-expressing HeLa cells. CONCLUSIONS The miR-720 promotes cell migration of HeLa cells by downregulating Rab35. The results show that miR-720 is a novel cell migration-associated gene in cervical cancer cells.

中文翻译：

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MicroRNA-720通过靶向Rab35在宫颈癌细胞中的表达来促进体外细胞迁移。

背景技术MicroRNA-720（miR-720）是一种非经典的miRNA，参与了几种肿瘤的发生和发展。在我们之前的研究中，与正常宫颈组织相比，miR-720在宫颈癌组织中被显着上调。但是，miR-720的确切生物学功能及其作用的分子机制仍然未知。结果微阵列表达谱，荧光素酶报告基因分析和蛋白质印迹分析被用于验证Rab35作为HEK293T和HeLa细胞中miR-720的靶基因。使用qRT-PCR和Western印迹分析评估了miR-720对Rab35表达的调节，并使用Transwell（®）分析，伤口愈合分析和real-in评估了外源性miR-720和Rab35对细胞迁移的影响。 HeLa细胞中的实时分析。通过HeLa细胞中的MTT测定，在体外评估了外源性miR-720对细胞增殖的影响。另外，在转染miR-720模拟物和Rab35表达载体后，还使用蛋白质印迹分析评估了与上皮-间质转化相关的E-钙粘蛋白和波形蛋白的表达。结果表明，小的GTPase Rab35是miR-720在宫颈癌HeLa细胞中的直接功能靶标。通过靶向Rab35，miR-720的过度表达导致E-钙黏着蛋白表达降低和波形蛋白表达增加，并最终导致HeLa细胞迁移的促进。此外，Rab35 3'-UTR（-）的重新引入显着逆转了表达miR-720的HeLa细胞中细胞迁移的诱导。结论miR-720通过下调Rab35促进HeLa细胞的细胞迁移。