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https://hdl.handle.net/2440/65593
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Type: | Journal article |
Title: | Selection of housekeeping genes for gene expression studies in a rat model of irinotecan-induced mucositis |
Author: | Al-Dasooqi, N. Bowen, J. Gibson, R. Logan, R. Stringer, A. Keefe, D. |
Citation: | Chemotherapy: international journal of experimental and clinical chemotherapy, 2011; 57(1):43-53 |
Publisher: | Karger |
Issue Date: | 2011 |
ISSN: | 0009-3157 1421-9794 |
Statement of Responsibility: | Noor Al-Dasooqi, Joanne M. Bowen, Rachel J. Gibson, Richard M. Logan, Andrea M. Stringer, Dorothy M. Keefe |
Abstract: | <h4>Background/aims</h4>Mucositis is the term used to describe damage caused by chemotherapy to mucous membranes of the alimentary tract. RT-PCR has recently been utilised to determine the molecular events that occur in mucositis. As this method relies on the use of a validated endogenous control, this study aims to validate commonly used housekeeping genes in an irinotecan-induced mucositis model.<h4>Methods</h4>Rats were administered irinotecan and sacrificed at different time points, in particular 1, 24, 72 and 144 h following treatment. Histopathological damage was assessed by haematoxylin and eosin staining. RT-PCR was used to evaluate the expression of 11 housekeeping genes. Expression stability was determined by the Normfinder program. Matrix metalloproteinase 2 was used as a target gene to validate the appropriateness of the top-ranking housekeeping gene.<h4>Results</h4>For normalisation to multiple housekeeping genes, the most stable combination across all time points in the jejunum was Ywhaz/UBC and in the colon UBC/β-actin. SDHA and GAPDH were the most variable genes in the jejunum and colon where they were 4.4 and 3.2 fold upregulated following irinotecan, respectively.<h4>Conclusions</h4>For normalisation of irinotecan-induced mucositis gene expression studies, a combination of Ywhaz/UBC and UBC/β-actin should be used in the jejunum and colon, respectively. UBC is the most favourable if restricted to a single housekeeping gene across all time points. |
Keywords: | Mucositis Alimentary tract Irinotecan RT-PCR Housekeeping genes |
Rights: | Copyright © 2011 S. Karger AG, Basel |
DOI: | 10.1159/000321477 |
Grant ID: | NHMRC |
Published version: | http://dx.doi.org/10.1159/000321477 |
Appears in Collections: | Aurora harvest 5 Medicine publications |
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