<Emphasis Type="Italic">RPN1</Emphasis>, a new reference gene for quantitative data normalization in lung and kidney cancer |
| |
Authors: | G S Krasnov N Yu Oparina A A Dmitriev A V Kudryavtseva E A Anedchenko T T Kondrat’eva E R Zabarovsky V N Senchenko |
| |
Institution: | 1.Engelhardt Institute of Molecular Biology,Russian Academy of Sciences,Moscow,Russia;2.Blokhin Cancer Research Center,Russian Academy of Medical Sciences,Moscow,Russia;3.MTC,Karolinska Institute,Stockholm,Sweden |
| |
Abstract: | Quantitative methods of gene expression analysis in tumors require accurate data normalization, which allows comparison of
different specimens with unknown mRNA/cDNA concentrations. For this purpose, reference genes with stable expression are used
(e.g., GAPDH, ACTB, HPRT1, or TBP). The problem of choosing proper reference genes is still a topical issue, because well-known reference genes can be unsuitable
for certain cancer types and their inappropriate use without additional testing can lead to wrong conclusions. A recently
developed bioinformatical approach was employed to identify a new potential reference gene for lung and kidney tumors, RPN1, located on the long arm of chromosome 3. The method employed the mining of the dbEST and Oncomine databases and functional
analysis of genes. RPN1 was selected from approximately 1500 candidate housekeeping genes. Using comparative genomic hybridization with NotI microarrays, we found no methylation, deletions, and/or amplifications in the RPN1-containing locus in 56 nonsmall cell lung and 42 clear cell renal cell cancer specimens. Real-time PCR showed that variation
of RPN1 mRNA levels in nonsmall cell lung cancer and clear-cell renal cancer was low and comparable to that of the known reference
genes GAPDH and GUSB, respectively. Expression levels of two hyalouronidase genes, HYAL1 and HYAL2, were assessed using the suggested references gene pairs (RPN1-GAPDH for lung cancer and RPN1-GUSB for kidney cancer), and these combinations were shown to produce accurate and reproducible data. These results suggest that
RPN1 is a new, promising reference gene for quantitative data normalization in gene expression studies for lung and kidney cancers. |
| |
Keywords: | |
本文献已被 SpringerLink 等数据库收录! |
|