Abstract | BACKGROUND/AIMS:
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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.
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Authors | Noor Al-Dasooqi, Joanne M Bowen, Rachel J Gibson, Richard M Logan, Andrea M Stringer, Dorothy M Keefe |
Journal | Chemotherapy
(Chemotherapy)
Vol. 57
Issue 1
Pg. 43-53
( 2011)
ISSN: 1421-9794 [Electronic] Switzerland |
PMID | 21282945
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Copyright | Copyright © 2011 S. Karger AG, Basel. |
Chemical References |
- 14-3-3 Proteins
- Actins
- Molecular Chaperones
- Ubiquitin C
- Ywhaz protein, rat
- Irinotecan
- Glyceraldehyde-3-Phosphate Dehydrogenases
- Succinate Dehydrogenase
- Matrix Metalloproteinase 2
- Camptothecin
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Topics |
- 14-3-3 Proteins
(genetics, metabolism)
- Actins
(genetics, metabolism)
- Animals
- Camptothecin
(analogs & derivatives, toxicity)
- Colon
(metabolism)
- Disease Models, Animal
- Gene Expression Regulation
(drug effects)
- Glyceraldehyde-3-Phosphate Dehydrogenases
(genetics, metabolism)
- Irinotecan
- Jejunum
(metabolism)
- Male
- Matrix Metalloproteinase 2
(genetics, metabolism)
- Molecular Chaperones
(genetics, metabolism)
- Mucositis
(chemically induced, genetics, metabolism)
- Rats
- Succinate Dehydrogenase
(genetics, metabolism)
- Ubiquitin C
(genetics, metabolism)
- Up-Regulation
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