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


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.


CONCLUSIONS
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.
Gerard
C J
CJ
diaDexus, LLC, Santa Clara, CA 95054, USA.
Andrejka
L M
LM
Macina
R A
RA
D003851:Deoxyribonucleases / Q000378:metabolism; D020869:Gene Expression Profiling; D006801:Humans; D008928:Mitochondria / Q000201:enzymology* / Q000235:genetics; D025261:Mitochondrial Proton-Translocating ATPases; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism*; D006180:Proton-Translocating ATPases / Q000032:analysis* / Q000235:genetics; D011786:Quality Control; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014018:Tissue Distribution
D003851:Deoxyribonucleases; C412204:ATP synthase subunit 6; D025261:Mitochondrial Proton-Translocating ATPases; D006180:Proton-Translocating ATPases