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Vol 47(2013) N 2 p. 192-196;
S.V. Fong1, S. Ibrahim2, M.S. Mohamed3, L.L. Few1, W.C.S. Too1, B.Y. Khoo4, Z. Yussof2, A.R.A. Rahman5, G.B. Yvonne-Tee1*

Identification of Endogenous Control Genes for Gene Expression Studies in Peripheral Blood of Patients with Coronary Artery Disease

1School of Health Sciences, University Sains Malaysia, Kubang Kerian, 16150, Malaysia
2School of Medical Sciences, University Sains Malaysia, Kubang Kerian, 16150, Malaysia
3Departament of Cardiology, Hospital Sultanah Nur Zahirah, Kuala Terengganu, 20400, Malaysia
4Institute for Research in Molecular Medicine (INFORMM), University Sains Malaysia, Penang, 11800, Malaysia
5Cyberjaya University College of Medical Sciences, Cyberjaya, 63000, Malaysia

*Yvonne@kb.usm.my
Received - 2012-07-26; Accepted - 2012-08-15

The selection of stable endogenous control genes is critical for normalization of quantitative realtime PCR (qPCR) data. In this study, we aimed to identify a suitable set of control genes to be used as endogenous references for gene expression evaluation in human peripheral blood samples among coronary artery disease patients. The expression levels of 12 endogenous control genes procured from TATAA Biocenter (Goteborg, Sweden) were measured in five acute coronary syndrome patients and five chronic stable angina patients. Gene expression stability was analyzed using two different software applications i.e geNorm and NormFinder. Results suggested that beta-glucuronidase is the most stable endogenous control, followed by hypoxanthine-guanine phosphoribosyltransferase. The NormFinder analysis further confirmed that beta-glucuronidase and hypoxanthine-guanine phosphoribosyltransferase were on the first rank order with the most stable expression among endogenous control genes analyzed and 60S acidic ribosomal protein P0. Besides, the expression levels of 18S rRNA were revealed to be highly variable between coronary heart disease patients. We thus recommend the use of beta-glucuronidase and hypoxanthine-guanine phosphoribosyltransferase as reference genes for accurate normalization of relative quantities of gene expression levels in coronary artery disease patients using qPCR. Also the use of 18S rRNA as a control gene should be avoided.

acute coronary syndrome, chronic stable angina, coronary artery disease, peripheral blood, endogenous control gene, gene expression



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