title: Integrated microRNA and mRNA Signature Associated with the Transition from the Locally Confined to the Metastasized Clear Cell Renal Cell Carcinoma Exemplified by miR-146-5p.
abstract: BACKGROUND
  MicroRNAs (miRNAs) regulate gene expression by interfering translation or stability of target transcripts. This interplay between miRNA and their mRNA has been proposed as an important process in cancer development and progression. We have investigated molecular networks impacted by predicted mRNA targets of differentially expressed miRNAs in patients with clear cell renal cell carcinoma (ccRCC) diagnosed with or without metastasis.
  
  
  METHODS
  miRNA and mRNA microarray expression profiles derived from primary ccRCC from patients with (16 samples) or without diagnosed metastasis (22 samples) were used to identify anti-correlated miRNA-mRNA interaction in ccRCC. For this purpose, Ingenuity pathway analysis microRNA Target Filter, which enables prioritization of experimentally validated and predicted mRNA targets was used. By applying an expression pairing tool, the analysis was focused on targets exhibiting altered expression in our analysis, finding miRNAs and their target genes with opposite or same expression. The resulting identified interactions were revalidated by RT-qPCR in another cohort of ccRCC patients. A selection of the predicted miRNA-mRNA interactions was tested by functional analyses using miRNA knockdown and overexpression experiments in renal cancer cell lines.
  
  
  RESULTS
  Among the significantly differentially expressed miRNAs, we have identified three miRNAs (miR-146a-5p, miR-128a-3p, and miR-17-5p) that were upregulated in primary tumors from patients without metastasis and downregulated in primary tumors from patients with metastasis. We have further identified mRNA targets, which expression were inversely correlated to these 3 miRNAs, and have been previously experimentally demonstrated in cancer setting in humans. Specifically, we showed that CXCL8/IL8, UHRF1, MCM10, and CDKN3 were downregulated and targeted by miR-146a-5p. The interaction between miR-146a-5p and their targets CXCL8 and UHRF1 was validated in cell culture experiments.
  
  
  CONCLUSIONS
  We identified novel target genes of dysregulated miRNAs, which are involved in the transition from primary RCC without metastases into tumors generating distant metastasis.
authors: Department of Urology, Charité - Universitätsmedizin Berlin, Berlin, Germany.|Berlin Institute for Urologic Research, Berlin, Germany.
authors: Zofia
authors: Z
authors: Wotschofsky
authors: Berlin Institute for Urologic Research, Berlin, Germany.|Department of General, Visceral, Vascular and Thoracic Surgery, Division of Molecular Biology, Charité - Universitätsmedizin Berlin, Berlin, Germany.
authors: Linda
authors: L
authors: Gummlich
authors: Department of Urology, Charité - Universitätsmedizin Berlin, Berlin, Germany.|Berlin Institute for Urologic Research, Berlin, Germany.
authors: Julia
authors: J
authors: Liep
authors: Department of Urology, Charité - Universitätsmedizin Berlin, Berlin, Germany.|Berlin Institute for Urologic Research, Berlin, Germany.
authors: Carsten
authors: C
authors: Stephan
authors: Institute of Pathology, Charité - Universitätsmedizin Berlin, Berlin, Germany.
authors: Ergin
authors: E
authors: Kilic
authors: Department of Urology, Charité - Universitätsmedizin Berlin, Berlin, Germany.|Berlin Institute for Urologic Research, Berlin, Germany.
authors: Klaus
authors: K
authors: Jung
authors: QIAGEN, 1700 Seaport Blvd, Redwood City, California, United States of America.
authors: Jean-Noel
authors: JN
authors: Billaud
authors: Department of Urology, Charité - Universitätsmedizin Berlin, Berlin, Germany.
authors: Hellmuth-Alexander
authors: HA
authors: Meyer
mesh_terms: D002292:Carcinoma, Renal Cell / Q000235:genetics* / Q000473:pathology; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007680:Kidney Neoplasms / Q000235:genetics* / Q000473:pathology; D035683:MicroRNAs / Q000378:metabolism / Q000502:physiology*; D009362:Neoplasm Metastasis / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: C542542:MIRN146 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger