title: MicroRNA-130b regulates the tumour suppressor RUNX3 in gastric cancer.
abstract: OBJECTIVE
  Accumulating evidence indicates that RUNX3 is an important tumour suppressor that is inactivated in many cancer types. This study aimed to assess the role of microRNA (miRNA) in the regulation of RUNX3.
  
  
  METHODS
  Four bioinformatic algorithms were used to predict miRNA binding to RUNX3. The correlation between candidate miRNAs and RUNX3 expression in cell lines was determined by real-time reverse transcriptase quantitative PCR (RT-qPCR) and Western blot. Candidate miRNAs were tested for functional effects through transfection of miRNA precursors and inhibitors, and monitoring cell viability, apoptosis and Bim expression. miRNA and RUNX3 expression, RUNX3 methylation and RUNX3 protein levels were assessed in gastric tissue by RT-qPCR, Methylight analysis and immunohistochemistry, respectively.
  
  
  RESULTS
  Bioinformatics, gene and protein expression analysis in eight gastric cell lines identified miR-130b as the top candidate miRNA for RUNX3 binding. Overexpression of miR-130b increased cell viability, reduced cell death and decreased expression of Bim in TGF-beta mediated apoptosis, subsequent to the downregulation of RUNX3 protein expression. In 15 gastric tumours, miR-130b expression was significantly higher compared to matched normal tissue, and was inversely associated with RUNX3 hypermethylation.
  
  
  CONCLUSIONS
  Attenuation of RUNX3 protein levels by miRNA may reduce the growth suppressive potential of RUNX3 and contribute to tumourigenesis.
authors.affiliation: Cancer Science Institute of Singapore, Centre for Life Sciences #02-15, 28 Medical Drive, National University of Singapore, Singapore 117456, Singapore.
authors.forename: Kin Wai
authors.initials: KW
authors.lastname: Lai
authors.forename: King Xin
authors.initials: KX
authors.lastname: Koh
authors.forename: Marie
authors.initials: M
authors.lastname: Loh
authors.forename: Kotaro
authors.initials: K
authors.lastname: Tada
authors.forename: Manish Mani
authors.initials: MM
authors.lastname: Subramaniam
authors.forename: Xn Yii
authors.initials: XY
authors.lastname: Lim
authors.forename: Aparna
authors.initials: A
authors.lastname: Vaithilingam
authors.forename: Manuel
authors.initials: M
authors.lastname: Salto-Tellez
authors.forename: Barry
authors.initials: B
authors.lastname: Iacopetta
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Ito
authors.forename: Richie
authors.initials: R
authors.lastname: Soong
mesh_terms: D000465:Algorithms; D045744:Cell Line, Tumor; D050677:Core Binding Factor Alpha 3 Subunit / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation / Q000502:physiology; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000235:genetics*; D016147:Genes, Tumor Suppressor; D006801:Humans; D007150:Immunohistochemistry; D035683:MicroRNAs / Q000378:metabolism*; D013274:Stomach Neoplasms / Q000235:genetics / Q000378:metabolism*
chemical_list: D050677:Core Binding Factor Alpha 3 Subunit; D035683:MicroRNAs; C493729:Runx3 protein, human