title: Targeting of RUNX3 by miR-130a and miR-495 cooperatively increases cell proliferation and tumor angiogenesis in gastric cancer cells.
abstract: Mature microRNAs (miRNAs) are 21 to 23 nucleotide noncoding RNA molecules that can downregulate multiple gene expression by mRNA degradation or translational repression. miRNAs are considered to play important roles in cell proliferation, apoptosis, and differentiation during mammalian development. The Runt-related transcription factor 3 (RUNX3) expression and activity are frequently downregulated by various mechanisms in gastric cancer. We have reported that RUNX3 inactivation is crucial for early tumorigenesis. In this study, we investigated the role of miRNAs targeting RUNX3 in early tumorigenesis. miR-130a and miR-495 upregulated under hypoxic conditions that bind to the RUNX3 3'-untranslated region (3'-UTR) were identified in gastric cancer cells by using microarray analysis and bioinformatics programs. Combination of miR-130a and miR-495 inhibited RUNX3 expression at the protein level, but not at the mRNA level. miR-130a and miR-495 significantly inhibited the RUNX3-3'UTR-luciferase activity. Combination of miR-130a and miR-495 significantly decreased apoptosis determined by Annexin V-FITC/propidium iodide staining and flow cytometric analysis, and the expression of Bim in SNU484 gastric cancer cells. In addition, p21 and Bim, RUNX3 target genes, were completely downregulated by the combination of miR-130a and miR-495. Using matrigel plug assay, we found that antagomiRs specific for miR-130a and miR-495 significantly reduced angiogenesis in vivo. In conclusion, targeting miR-130a and miR-495 could be a potential therapeutics to recover RUNX3 expression under hypoxic conditions and in early tumorigenic progression.
authors.affiliation: Research Institute of Pharmaceutical Sciences, College of Pharmacy, Daegu, 702-701, Republic of Korea.
authors.forename: Sun Hee
authors.initials: SH
authors.lastname: Lee
authors.affiliation: Research Institute of Pharmaceutical Sciences, College of Pharmacy, Daegu, 702-701, Republic of Korea.
authors.forename: Yuk Dong
authors.initials: YD
authors.lastname: Jung
authors.affiliation: Research Institute of Pharmaceutical Sciences, College of Pharmacy, Daegu, 702-701, Republic of Korea.
authors.forename: Young Sun
authors.initials: YS
authors.lastname: Choi
authors.affiliation: Research Institute of Pharmaceutical Sciences, College of Pharmacy, Daegu, 702-701, Republic of Korea.
authors.forename: You Mie
authors.initials: YM
authors.lastname: Lee
mesh_terms: D049109:Cell Proliferation / Q000235:genetics*; D050677:Core Binding Factor Alpha 3 Subunit / Q000235:genetics*; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D057809:HEK293 Cells; D006801:Humans; D035683:MicroRNAs / Q000502:physiology*; D046228:Microarray Analysis; D009389:Neovascularization, Pathologic / Q000235:genetics*; D013274:Stomach Neoplasms / Q000098:blood supply / Q000235:genetics* / Q000473:pathology; D014407:Tumor Cells, Cultured
chemical_list: D050677:Core Binding Factor Alpha 3 Subunit; C554019:MIRN130 microRNA, human; C577768:MIRN495 microRNA, human; D035683:MicroRNAs; C493729:Runx3 protein, human
keywords: RUNX3; angiogenesis; cell proliferation; miR-130a; miR-495