The miR-17-92 Cluster is Over Expressed in and Has an Oncogenic Effect on Renal Cell Carcinoma
Received 29 April 2009 published online 17 December 2009.
Purpose
miRNAs are small, nonprotein coding RNAs that are differentially expressed in many malignancies. We previously identified 80 miRNAs that are dysregulated in clear cell renal cell carcinoma. In this study we validated over expression of the miR-17-92 cluster in clear cell renal cell carcinoma and tested the effect of 2 members of this cluster (miR-17-5p and miR-20a) on tumor proliferation. We also elucidated the role of miRNA in clear cell renal cell carcinoma pathogenesis with bioinformatics.
Materials and Methods
miRNA expression was validated by quantitative reverse transcriptase-polymerase chain reaction. The cell proliferation effect of miR-17-5p and miR-20a was tested in a renal adenocarcinoma cell line model. Multiple in silico analyses were done of dysregulated miRNAs.
Results
We validated miR-71-92 cluster over expression in clear cell renal cell carcinoma by quantitative reverse transcriptase-polymerase chain reaction. Transfection of miR-20a inhibitor significantly decreased cell proliferation in a dose dependent manner. Transfection of miR-17-5p, which is not endogenously expressed in the ACHN cell line, led to increased cell proliferation compared to control values. This effect was suppressed by miR-17-5p inhibitor. Bioinformatics analysis identified 10 clusters of miRNAs dysregulated in clear cell renal cell carcinoma that followed the same expression patterns. We also identified matching patterns between reported chromosomal aberration in clear cell renal cell carcinoma and miRNA dysregulation for 37.5% of the miRNAs. Target prediction analysis was done using multiple algorithms. Many key molecules in clear cell renal cell carcinoma pathogenesis, including HIFs, mTOR, VEGF and VHL, were potential targets for dysregulated miRNAs.
Conclusions
A significant number of dysregulated proteins in clear cell renal cell carcinoma are potential miRNA targets. Also, many clear cell renal cell carcinoma dysregulated miRNAs are phylogenetically conserved.
aDepartment of Laboratory Medicine and Keenan Research Centre, Li Ka Shing Knowledge Institute, Toronto, Ontario, Canada
bDivision of Urology, Department of Surgery, St. Michael's Hospital, Toronto, Ontario, Canada
cDepartment of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, Canada
dDepartment of Biochemistry and Molecular Biology, Faculty of Biology, University of Athens, Panepistimiopolis, Athens, Greece
Correspondence and requests for reprints: Department of Laboratory Medicine, St. Michael's Hospital, 30 Bond St., Toronto, Ontario, M5B 1W8, Canada (telephone: 416-864-6060, extension 6129; FAX: 416-864-5648)
Study received approval from the research ethics board, St. Michael's Hospital, Toronto, Canada.
Supported by Canadian Institute of Health Research Grant 86490.
ABSTRACT