Selection of housekeeping genes for real-time PCR in atopic human bronchial epithelial cells
The stability of housekeeping genes (HKGs) is critical when performing real-time quantitative PCR. To date, the stability of common HKGs has not been systematically compared in human airway epithelial cells (AEC) in normal and atopic subjects. Expression levels of 12 HKGs were measured in AECs from...
| Main Authors: | , , , , , , , |
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| Format: | Journal Article |
| Language: | English |
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EUROPEAN RESPIRATORY SOC JOURNALS LTD
2008
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| Online Access: | http://hdl.handle.net/20.500.11937/76826 |
| _version_ | 1848763770199867392 |
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| author | He, J.Q. Sandford, A.J. Wang, I.M. Stepaniants, S. Knight, D.A. Kicic, Anthony Stick, S.M. Paré, P.D. |
| author_facet | He, J.Q. Sandford, A.J. Wang, I.M. Stepaniants, S. Knight, D.A. Kicic, Anthony Stick, S.M. Paré, P.D. |
| author_sort | He, J.Q. |
| building | Curtin Institutional Repository |
| collection | Online Access |
| description | The stability of housekeeping genes (HKGs) is critical when performing real-time quantitative PCR. To date, the stability of common HKGs has not been systematically compared in human airway epithelial cells (AEC) in normal and atopic subjects. Expression levels of 12 HKGs were measured in AECs from a cohort of 30 healthy atopic nonasthmatic or atopic asthmatic children. Gene expression stability was determined using three different Visual Basic for Applications applets (geNorm, NormFinder and BestKeeper). All 12 HKGs were expressed in AECs. However, the hypoxanthine ribosyltransferase and TATA-binding protein genes were excluded from further analysis due to low expression levels. The cyclophilin A gene was ranked the most stable by all three methods. The expression levels of the β-actin and glyceraldehyde-3-phosphate dehydrogenase genes were significantly different between the three groups of patients, with atopic asthmatics showing the highest expression levels for both genes. The results suggest that the cyclophilin A gene is the most suitable housekeeping gene analysed for expression studies utilising uncultured bronchial airway epithelial cells from healthy and asthmatic children, and highlight the importance of validating housekeeping genes for each experimental model. Copyright©ERS Journals Ltd 2008. |
| first_indexed | 2025-11-14T11:08:44Z |
| format | Journal Article |
| id | curtin-20.500.11937-76826 |
| institution | Curtin University Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-14T11:08:44Z |
| publishDate | 2008 |
| publisher | EUROPEAN RESPIRATORY SOC JOURNALS LTD |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | curtin-20.500.11937-768262019-11-12T08:19:30Z Selection of housekeeping genes for real-time PCR in atopic human bronchial epithelial cells He, J.Q. Sandford, A.J. Wang, I.M. Stepaniants, S. Knight, D.A. Kicic, Anthony Stick, S.M. Paré, P.D. Science & Technology Life Sciences & Biomedicine Respiratory System atopic asthma atopy housekeeping genes human bronchial epithelial cells real-time quantitative PCR RT-PCR BETA-ACTIN EXPRESSION AIRWAY NORMALIZATION ASTHMA CYCLOPHILIN CYTOKINE INJURY The stability of housekeeping genes (HKGs) is critical when performing real-time quantitative PCR. To date, the stability of common HKGs has not been systematically compared in human airway epithelial cells (AEC) in normal and atopic subjects. Expression levels of 12 HKGs were measured in AECs from a cohort of 30 healthy atopic nonasthmatic or atopic asthmatic children. Gene expression stability was determined using three different Visual Basic for Applications applets (geNorm, NormFinder and BestKeeper). All 12 HKGs were expressed in AECs. However, the hypoxanthine ribosyltransferase and TATA-binding protein genes were excluded from further analysis due to low expression levels. The cyclophilin A gene was ranked the most stable by all three methods. The expression levels of the β-actin and glyceraldehyde-3-phosphate dehydrogenase genes were significantly different between the three groups of patients, with atopic asthmatics showing the highest expression levels for both genes. The results suggest that the cyclophilin A gene is the most suitable housekeeping gene analysed for expression studies utilising uncultured bronchial airway epithelial cells from healthy and asthmatic children, and highlight the importance of validating housekeeping genes for each experimental model. Copyright©ERS Journals Ltd 2008. 2008 Journal Article http://hdl.handle.net/20.500.11937/76826 10.1183/09031936.00129107 English EUROPEAN RESPIRATORY SOC JOURNALS LTD restricted |
| spellingShingle | Science & Technology Life Sciences & Biomedicine Respiratory System atopic asthma atopy housekeeping genes human bronchial epithelial cells real-time quantitative PCR RT-PCR BETA-ACTIN EXPRESSION AIRWAY NORMALIZATION ASTHMA CYCLOPHILIN CYTOKINE INJURY He, J.Q. Sandford, A.J. Wang, I.M. Stepaniants, S. Knight, D.A. Kicic, Anthony Stick, S.M. Paré, P.D. Selection of housekeeping genes for real-time PCR in atopic human bronchial epithelial cells |
| title | Selection of housekeeping genes for real-time PCR in atopic human bronchial epithelial cells |
| title_full | Selection of housekeeping genes for real-time PCR in atopic human bronchial epithelial cells |
| title_fullStr | Selection of housekeeping genes for real-time PCR in atopic human bronchial epithelial cells |
| title_full_unstemmed | Selection of housekeeping genes for real-time PCR in atopic human bronchial epithelial cells |
| title_short | Selection of housekeeping genes for real-time PCR in atopic human bronchial epithelial cells |
| title_sort | selection of housekeeping genes for real-time pcr in atopic human bronchial epithelial cells |
| topic | Science & Technology Life Sciences & Biomedicine Respiratory System atopic asthma atopy housekeeping genes human bronchial epithelial cells real-time quantitative PCR RT-PCR BETA-ACTIN EXPRESSION AIRWAY NORMALIZATION ASTHMA CYCLOPHILIN CYTOKINE INJURY |
| url | http://hdl.handle.net/20.500.11937/76826 |