Mitochondrial ATP synthase 6 as an endogenous control in the quantitative RT-PCR analysis of clinical cancer samples

Real-time polymerase chain reaction (PCR) is a powerful new technique in the evolution of quantitative reverse transcription-PCR assays. With the increased sensitivity and resolution of real-time techniques, the requirements for constitutive expression of endogenous controls have become increasingly...

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Bibliographic Details
Published in:Molecular diagnosis 2000-03, Vol.5 (1), p.39-46
Main Authors: Gerard, C J, Andrejka, L M, Macina, R A
Format: Article
Language:English
Subjects:
Deoxyribonucleases - metabolism
Gene Expression Profiling
Humans
Mitochondria - enzymology
Mitochondria - genetics
Mitochondrial Proton-Translocating ATPases
Neoplasms - genetics
Neoplasms - metabolism
Proton-Translocating ATPases - analysis
Proton-Translocating ATPases - genetics
Quality Control
Reference Standards
Reverse Transcriptase Polymerase Chain Reaction - methods
Tissue Distribution
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Summary:Real-time polymerase chain reaction (PCR) is a powerful new technique in the evolution of quantitative reverse transcription-PCR assays. With the increased sensitivity and resolution of real-time techniques, the requirements for constitutive expression of endogenous controls have become increasingly stringent. We compare the expression of the mitochondrial gene, adenosine triphosphate synthase 6 (ATPsy6), to the expression of other routinely used endogenous control genes (e.g., beta-actin, glyceraldehyde-3-phosphate dehydrogenase [GAPDH], ribosomal RNA 18S [18S rRNA], and cyclophilin). In a diverse assortment of tissues and across a wide range of disease stages, ATPsy6 shows a relative steady state of expression compared with other endogenous controls. ATPsy6 gene expression has been used as an endogenous control in a quantitative real-time PCR assay designed to evaluate the expression of potential cancer diagnostic leads across a diverse tissue panel. Mitochondrial ATPsy6 serves as a good endogenous control to measure target gene expression independent of the tissue- or disease-specific variation inherent with many housekeeping genes.
ISSN:1084-8592
DOI:10.1007/BF03262021