Abstract
MicroRNAs (miRNAs) are endogenous small non-coding RNAs that play central roles in diverse pathological processes. In this study, we investigated the effect of microRNA-182 (miR-182) on the development of posterior uveal melanomas. Initially, we demonstrated that miR-182 expression was dependent on p53 induction in uveal melanoma cells. Interestingly, transient transfection of miR-182 into cultured uveal melanoma cells led to a significant decrease in cell growth, migration, and invasiveness. Cells transfected with miR-182 demonstrated cell cycle G1 arrest and increased apoptotic activity. Using bioinformatics, we identified three potential targets of miR-182, namely MITF, BCL2 and cyclin D2. miR-182 was shown to have activity on mRNA expression by targeting the 3' untranslated region of MITF, BCL2 and cyclin D2. Subsequent Western blot analysis confirmed the downregulation of MITF, BCL2 and cyclin D2 protein expression. The expression of oncogene c-Met and its downstream Akt and ERK1/2 pathways was also downregulated by miR-182. Concordant with the findings that miR-182 was decreased in uveal melanoma tissue samples, overexpression of miR-182 also suppressed the in vivo growth of uveal melanoma cells. Our results demonstrated that miR-182, a p53 dependent miRNA, suppressed the expression of MITF, BCL2, cyclin D2 and functioned as a potent tumor suppressor in uveal melanoma cells.
Publication types
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Clinical Trial
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Research Support, Non-U.S. Gov't
MeSH terms
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3' Untranslated Regions / genetics
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Cell Line, Tumor
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Cyclin D2 / biosynthesis*
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Cyclin D2 / genetics
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Down-Regulation*
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Female
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Gene Expression Regulation, Neoplastic*
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Genes, Tumor Suppressor*
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Humans
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MAP Kinase Signaling System / genetics
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Male
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Melanoma / genetics
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Melanoma / metabolism*
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Melanoma / pathology
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MicroRNAs / biosynthesis*
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MicroRNAs / genetics
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Microphthalmia-Associated Transcription Factor / biosynthesis*
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Microphthalmia-Associated Transcription Factor / genetics
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Mitogen-Activated Protein Kinase 1 / genetics
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Mitogen-Activated Protein Kinase 1 / metabolism
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Mitogen-Activated Protein Kinase 3 / genetics
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Mitogen-Activated Protein Kinase 3 / metabolism
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Proto-Oncogene Proteins c-akt / genetics
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Proto-Oncogene Proteins c-akt / metabolism
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Proto-Oncogene Proteins c-bcl-2 / biosynthesis*
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Proto-Oncogene Proteins c-bcl-2 / genetics
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RNA, Neoplasm / biosynthesis*
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RNA, Neoplasm / genetics
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Tumor Suppressor Protein p53 / genetics
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Tumor Suppressor Protein p53 / metabolism
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Uveal Neoplasms / genetics
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Uveal Neoplasms / metabolism*
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Uveal Neoplasms / pathology
Substances
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3' Untranslated Regions
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CCND2 protein, human
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Cyclin D2
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MITF protein, human
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MicroRNAs
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Microphthalmia-Associated Transcription Factor
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Mirn182 microRNA, human
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Proto-Oncogene Proteins c-bcl-2
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RNA, Neoplasm
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TP53 protein, human
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Tumor Suppressor Protein p53
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Proto-Oncogene Proteins c-akt
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MAPK1 protein, human
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Mitogen-Activated Protein Kinase 1
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Mitogen-Activated Protein Kinase 3
Grants and funding
This work was supported, in part, by the National Natural Science Foundation of China Grant 81071682, Zhejiang Provincial Natural Science Foundation of China Grant Y2080853, Program for New Century Excellent Talents in University (NCET-06-0538), 973 Projects (2011CB504605 & 2012CB22303) from the Ministry of Science and Technology of China, Major Program of Science Foundation of the Affiliated Eye Hospital of Wenzhou Medical College YNZD201002 and Science Foundation of Wenzhou Medical College QTJ11020. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. No additional external funding was received for this study.