Abstract
Purpose: :
MicroRNAs (miRNAs) are endogenous 20-24 nucleotide RNAs which inhibit protein translation through binding to target mRNAs. Recent studies have demonstrated that miRNAs played important roles in tumorigenesis. The role of miRNAs in uveal melanoma, however, remains largely unknown. In the present study, we investigated the function of (MicroRNA-96) miR-96 in uveal melanoma cells.
Methods: :
Realtime-PCR was performed to detect the expression of miR-96 in uveal melanoma cells. The proliferation of uveal melanoma cells was quantified by MTS assay. The target of miR-96 was predicted by bioinformatics and confirmed by luciferase assay. Uveal melanoma cells were transfected with miR-96 by liperfectamine 2000. The expression of MITF was analyzed by Western blotting.
Results: :
miR-96 was upregulated in uveal melanoma cells after treated with adriamycin. Introduction of miR-96 into uveal melanoma cells resulted in a significant decrease in cell growth. Four putative miR-96 binding sites were predicted within the 3’ untranslated region (3’ UTR) of human MITF 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-96. miR-96 was found to downregulate the expression of MITF by Western blot analysis.
Conclusions: :
Our results demonstrated that miR-96 may act as a tumor suppressor in uveal melanoma cell proliferation through downregulation of MITF.
Keywords: melanoma • uvea • proliferation