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Identification of novel dioxin-responsive genes by representational difference analysis
S P Rivera1, S T Saarikoski, W Sun
1Department of Pathology and Laboratory Medicine, and Jonsson Comprehensive Cancer Center, University of California, Los Angeles, CA, USA.
Abstract:
Representational difference analysis (RDA) was employed on dioxin-stimulated Hepa-1 mouse hepatoma cells (human breast cancer) MCF-7 cells, mouse liver, and mouse thymocytes in order to identify novel responder genes to dioxin. In addition to several clones representing known dioxin-inducible genes, several clones were isolated that represented genes that were previously not known to be inducible by dioxin, including B94 (also known as tumour necrosis factor alpha-induced protein 2 (Tnfaip2), Seven in absentia homologue 2 (Siah2), the mouse homologue of Bob/Gpr15, and S-adenosyl-homocysteine hydrolase (Sahh, Ahcy). Induction of these genes by dioxin in Hepa-1 cells was rapid. Furthermore, induction occurred in the presence of the protein synthesis inhibitor cycloheximide, indicating that in each case induction is a primary response.
Insights
This study identified novel dioxin-responsive genes in mouse cells using representational difference analysis (RDA). Several previously unknown dioxin-inducible genes were discovered, indicating a primary response mechanism.
Area of Science:
- Molecular Biology
- Toxicology
- Genetics
Background:
- Dioxins are environmental pollutants known to induce gene expression.
- Identifying dioxin-responsive genes is crucial for understanding their toxicological effects.
- Previous studies have identified some dioxin-inducible genes, but a comprehensive understanding is lacking.
Purpose of the Study:
- To identify novel genes that are rapidly induced by dioxin.
- To investigate the primary response of these novel genes to dioxin exposure.
- To utilize representational difference analysis (RDA) for novel gene discovery.
Main Methods:
- Representational difference analysis (RDA) was performed on dioxin-stimulated Hepa-1 mouse hepatoma cells, MCF-7 cells, mouse liver, and mouse thymocytes.
- Differential gene expression was analyzed to identify novel dioxin-inducible genes.
- The induction of identified genes was confirmed in Hepa-1 cells, including in the presence of cycloheximide.
Main Results:
- Several known dioxin-inducible genes were identified.
- Novel dioxin-inducible genes were discovered, including B94 (Tnfaip2), Siah2, the mouse homologue of Bob/Gpr15, and Sahh (Ahcy).
- Dioxin induction of these novel genes in Hepa-1 cells was rapid and occurred even with cycloheximide, suggesting a primary response.
Conclusions:
- Representational difference analysis (RDA) is effective in identifying novel dioxin-responsive genes.
- The identified genes (B94, Siah2, Bob/Gpr15 homologue, Sahh) represent new targets for dioxin toxicity research.
- Dioxin induction of these genes is a primary response, independent of new protein synthesis.
