PURPOSE: Real-time quantitative PCR is a very powerful technique to examine gene expression profiles under different biological conditions. The stability of housekeeping genes is critical when comparing gene expression profiles in normal and disease tissues. To date, there have been no studies that have no studies which have systematically compared the stability of common housekeeping genes in normal and atopic human bronchial epithelial cells.
METHODS: We measured the expression levels of twelve housekeeping genes and analyzed the stability of their expression in bronchial epithelial cells of healthy children (n = 10), atopic children without asthma (n = 10) and atopic asthmatic children (n = 10). The twelve housekeeping genes studied were 18S rRNA, Acidic ribosomal protein (PO), Beta-actin (BA), Cyclophilin (CYC), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Phosphoglycerokinase (PGK), B-2-Microglobulin (B2M), Glucronidase (GUS), Hypoxanthine ribosyl transferase (HPRT), Transcription factor IID, TATA binding protein (TBP), Transferrin receptor (TfR) and Guanine nucleotide binding protein (GNB2L1).
RESULTS: All twelve housekeeping genes were expressed in human airway epithelial cells. However, two of these genes (HPRT and TBP) were excluded from stability analysis due to relative low expression levels. The stability of the remaining ten genes was determined using geNorm applet. The three most stable genes were CYC, GNB2L1 and 18S rNA in healthy children; GNB2L1, BA and CYC in atopic children without asthma; BA, GAPDH and CYC in atopic asthmatic children.
CONCLUSION: The results of this study suggest that CYC may be the most suitable housekeeping gene in gene expression studies of human bronchial epithelial cells derived from normal and atopic children.
CLINICAL IMPLICATIONS: This study provides important information for gene expression studies of asthmatic bronchial epithelial cell - a target to identify novel mechanisms of asthma pathogenesis and new therapeutic strategies.
DISCLOSURE: Jian-Qing He, No Financial Disclosure Information; No Product/Research Disclosure Information