NDLI: RPN1, a new reference gene for quantitative data normalization in lung and kidney cancer

Content Provider	Springer Nature Link
Author	Krasv, G. S. Oparina, N. Yu. Dmitriev, A. A. Kudryavtseva, A. V. Anedchenko, E. A. Kondrat’eva, T. T. Zabarovsky, E. R. Senchenko, V. N.
Copyright Year	2011
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.
Starting Page	211
Ending Page	220
Page Count	10
File Format	PDF
ISSN	00268933
Journal	Molecular Biology
Volume Number	45
Issue Number	2
e-ISSN	16083245
Language	English
Publisher	SP MAIK Nauka/Interperiodica
Publisher Date	2011-04-26
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	reference genes normalization ribophorin nonsmall cell lung cancer clear cell renal cancer Human Genetics Biochemistry Life Sciences
Content Type	Text
Resource Type	Article
Subject	Structural Biology Biophysics

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