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

Abstract

BACKGROUND:

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.

METHODS AND RESULTS:

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.

CONCLUSION:

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.

• Authors: C J Gerard, L M Andrejka, R A Macina
• Journal: Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology (Mol Diagn) Vol. 5 Issue 1 Pg. 39-46 (Mar 2000) ISSN: 1084-8592 [Print] United States
• PMID: 10837088 (Publication Type: Journal Article)

Chemical References

• Deoxyribonucleases
• ATP synthase subunit 6
• Mitochondrial Proton-Translocating ATPases
• Proton-Translocating ATPases

Topics

• Deoxyribonucleases (metabolism)
• Gene Expression Profiling
• Humans
• Mitochondria (enzymology, genetics)
• Mitochondrial Proton-Translocating ATPases
• Neoplasms (genetics, metabolism)
• Proton-Translocating ATPases (analysis, genetics)
• Quality Control
• Reference Standards
• Reverse Transcriptase Polymerase Chain Reaction (methods)
• Tissue Distribution