Abstract
BackgroundMicroRNA-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.ResultsMicroarray 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.ConclusionsThe 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.Electronic supplementary materialThe online version of this article (doi:10.1186/s13578-015-0047-5) contains supplementary material, which is available to authorized users.
Highlights
MicroRNA-720, a nonclassical miRNA, is involved in the initiation and progression of several tumors
Some studies showed that Wnt5a promotes breast cancer cell migration via the Dvl2/Rab35/Rac1 signaling pathway [36], and Rab35 maintains cadherins at the cell surface to promote cell–cell adhesion [37]
Results miR‐720 promotes cell migration but does not affect cell proliferation in HeLa cells In our previous study, we found the expression level of miR-720 was significantly upregulated in cervical cancer tissues compared to normal adjacent tissues [17]
Summary
MicroRNA-720 (miR-720), a nonclassical miRNA, is involved in the initiation and progression of several tumors. MiR-720 was shown to be significantly upregulated in cervical cancer tissues compared with normal cervical tissues. Discovered in 1993, microRNAs (miRNAs) are a class of short, non-coding RNAs that are highly efficient gene expression regulators in various cellular processes [1,2,3,4,5]. They modulate gene expression predominantly through an interaction with the 3′-UTR of their target mRNAs [6, 7]. The roles of miR-720 in the initiation and progression of cervical cancer are still largely unknown. Some studies showed that Wnt5a promotes breast cancer cell migration via the Dvl2/Rab35/Rac signaling pathway [36], and Rab maintains cadherins at the cell surface to promote cell–cell adhesion [37]
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