miR-128 targets the CC chemokine ligand 18 gene (CCL18) in cutaneous malignant melanoma progression

摘要

BackgroundThe CC chemokine ligand 18 (CCL18) has a higher expression in some tumors, while the CCL18 level can be a marker of tumor progression and prognosis. We previously reported that the expression ofCCL18gene was dramatically up-regulated in cutaneous malignant melanoma (CMM) and its expression levels were correlated with tumor thickness. ObjectiveTo investigate miRNAs which could target theCCL18gene so as to mediate CMM development and improvement. MethodsThe expression of miR-128 andCCL18in CMM were measured by qRT-PCR. The interaction of miR-128 withCCL183′UTR was verified by Luciferase reporter gene assay. The changes in expression ofCCL18after miR-128 mimic transfection of A375 melanoma cells were determined by both qRT-PCR and Western-bloting. Cell viability was accessed by CCK8-assay. Flow cytometry was employed to detect the incidence of apoptosis. Clonogenic assay was used to detect the ability of colony formation. Cell migration was evaluated by Transwell migration study. The protein levels of epithelial-mesenchymal transition (EMT), such as E-cadherin, N-cadherin and β-catenin were analyzed by Western-bloting. ResultsThe expression of miR-128 had negative relevance withCCL18in CMM. miR-128 could interact withCCL183′UTR. Transfected miR-128 mimic significantly reducedCCL18expression and this impairment ofCCL18gene promoted apoptosis, inhibited migration and colony formation of A375 melanoma cells. Furthermore, the relative expression of N-cadherin was decreased. ConclusionCCL18is a target gene of miR-128. Overexpression of miR-128 inhibits the oncogenic effect ofCCL18.