Gene expression analysis of the rat testis after treatment with di(2-ethylhexyl) phthalate using cDNA microarray and real-time RT-PCR
| Autor: | Masatoshi Komiyama, Tetsuya Adachi, Kazuyasu Kijima, Kaoru Toyosawa, Masashi Yasuba, Chisato Mori, Nobuo Matsuoka |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Male
Programmed cell death Fas Ligand Protein Apoptosis Toxicology Fas ligand Rats Sprague-Dawley Random Allocation chemistry.chemical_compound Plasticizers Diethylhexyl Phthalate Testis Gene expression In Situ Nick-End Labeling Animals RNA Messenger fas Receptor FADD Oligonucleotide Array Sequence Analysis bcl-2-Associated X Protein Pharmacology Membrane Glycoproteins TUNEL assay biology Histocytochemistry Reverse Transcriptase Polymerase Chain Reaction Phthalate Molecular biology Rats Isoenzymes Reverse transcription polymerase chain reaction Real-time polymerase chain reaction Gene Expression Regulation Proto-Oncogene Proteins c-bcl-2 chemistry Caspases biology.protein |
| Zdroj: | Toxicology and Applied Pharmacology. 200:103-110 |
| ISSN: | 0041-008X |
| Popis: | To investigate the effects of di(2-ethylhexyl) phthalate (DEHP) on gene expression in rat testis, 6-week-old male Sprague-Dawley rats were given a single oral dose of 20 or 2000 mg/kg and euthanized 3, 6, 24, or 72 h thereafter. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive cells were significantly increased in the testis at 24 and 72 h after the exposure to 2000 mg/kg of DEHP. On cDNA microarray analysis, in addition to apoptosis-related genes, genes associated with atrophy, APEX nuclease, MutS homologue (E. coli), testosterone-repressed-prostatic-message-2 (TRPM-2), connective tissue growth factor, collagen alpha 2 type V, and cell adhesion kinase were differentially expressed. To investigate the relationship between histopathological alteration and gene expression, we selected genes associated with apoptosis and analyzed their expression by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). With 20 mg/kg of DEHP treatment, bcl-2, key gene related to apoptosis, was increased. Up-regulation of bcl-2, inhibitor of Apaf-1/caspase-9/caspase-2 cascade of apoptosis, may be related to the fact that no morphological apoptotic change was induced after dosing of 20 mg/kg DEHP. With 2000 mg/kg of DEHP treatment, the apoptotic activator cascade, Fas/FasL, FADD/caspase-8/caspase-3 cascade, and Apaf-1/caspase-9/caspase-2 cascade were increased and bcl-2 was decreased. Thus, these gene regulations might lead the cells into apoptosis in the case of high exposure to DEHP. In contrast, FADD/caspase-10/caspase-6 cascade and caspase-11/caspase-3 cascade were not increased. These results indicate that the cascades of FADD/caspase-10/caspase-6 and caspase-11/caspase-3 are not related to apoptosis with DEHP treatment. |
| Databáze: | OpenAIRE |
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