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MicroRNA 145 may play an important role in uveal melanoma cell growth by potentially targeting insulin receptor substrate-1 / 中华医学杂志(英文版)
Chinese Medical Journal ; (24): 1410-1416, 2014.
Article in English | WPRIM | ID: wpr-322256
Responsible library: WPRO
ABSTRACT
<p><b>BACKGROUND</b>MicroRNAs (miRNAs) contribute to tumorigenesis by acting as either oncogenes or tumor suppressor genes. In this study, we investigated the role of miR-145 in the pathogenesis of uveal melanoma.</p><p><b>METHODS</b>Expression profiles of miRNAs in uveal melanoma were performed using Agilent miRNA array. Quantitative real-time polymerase chain reaction was used to screen the expression levels of miR-145 in normal uveal tissue, uveal melanoma tissue, and uveal melanoma cell lines. Lenti-virus expression system was used to construct MUM-2B and OCM-1 cell lines with stable overexpression of miR-145. Cell proliferation, cell cycle, and cell apoptosis of these miR-145 overexpression cell lines were examined by MTT assay and flow cytometry respectively. The target genes of miR-145 were predicted by bioinformatics and confirmed using a luciferase reporter assay. The expression of insulin-like growth factor-1 receptor (IGF-1R), insulin receptor substrate-1 (IRS-1) proteins was determined by Western blotting analysis. IRS-1 was knocked down in OCM-1 cells. TUNEL, BrdU, and flow cytometry assay were performed in IRS-1 knocked down OCM-1 cell lines to analyze its function.</p><p><b>RESULTS</b>Forty-seven miRNAs were up regulated in uveal melanoma and 61 were down regulated. miR-145 expression was significantly lower in uveal melanoma sample and the cell lines were compared with normal uveal sample. Overexpression of miR-145 suppressed cell proliferation by blocking the G1 phase entering S phase in uveal melanoma cells, and promoted uveal melanoma cell apoptosis. IRS-1 was identified as a potential target of miR-145 by dual luciferase reporter assay. Knocking down of IRS-1 had similar effect as overexpression of miR-145.</p><p><b>CONCLUSION</b>miR-145 might act as a tumor suppressor in uveal melanoma, and downregulation of the target IRS-1 might be a potential mechanism.</p>
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Pathology / Physiology / Uveal Neoplasms / In Vitro Techniques / Humans / Cell Cycle / Blotting, Western / Polymerase Chain Reaction / Apoptosis / MicroRNAs Language: English Journal: Chinese Medical Journal Year: 2014 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Pathology / Physiology / Uveal Neoplasms / In Vitro Techniques / Humans / Cell Cycle / Blotting, Western / Polymerase Chain Reaction / Apoptosis / MicroRNAs Language: English Journal: Chinese Medical Journal Year: 2014 Type: Article