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D. Yan, X. Zhou, D.-N. Hu, X. Chen, J. Wang, F. Lu, L. Tu, J. Qu; Microrna-34a Downregulates C-met Expression in Uveal Melanoma Cells. Invest. Ophthalmol. Vis. Sci. 2008;49(13):4444.
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MicroRNAs (miRNAs) are endogenously expressed 20-24 nucleotide RNAs which repress protein translation through binding to a target mRNA. Recent studies have shown that some miRNAs can regulate proliferation of tumor cells. The role of miRNAs in uveal melanoma, however, remains unclear. In the present study, we investigated the function of miR-34a in uveal melanoma cells.
Northern blotting was performed to detect the expression of miR-34a in melanoma cells and melanocytes. The target of miR-34a was predicted by bioinformatics and confirmed by luciferase assay. Uveal melanoma cells were transfected with miR-34a. The amount of c-met protein was determined by Western blotting to evaluate the effect of miR-34a on c-met. The proliferation of melanoma cells was quantified by MTT assay.
miR-34a was expressed in melanocytes but not in uveal melanoma cells. Two putative miR-34a binding sites were found in silico within the 3’ untranslated region (3’ UTR) of human c-met mRNA. Reporter constructs containing the binding sites inserted into the 3’ UTR of the luciferase mRNA conferred a repression of luciferase activity in HEK293 cells transfected with miR-34a. miR-34a was found to downregulate the expression of c-met protein by Western blot analysis. Transfection of miR-34a into uveal melanoma cells led to a significant decrease in cell growth.
Our results demonstrated that miR-34a was involved in uveal melanoma cell proliferation by targeting c-met. This indicates that miR-34a may act as a suppressor of uveal melanoma cell proliferation and growth.
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