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Identification of the genes responsive to etoposide-induced apoptosis: application of DNA chip technology
1Department of Molecular Biology, Parke-Davis Pharmaceutical Research, Division of Warner-Lambert Company, Ann Arbor, MI 48105, USA.
Abstract:
DNA chip technology was used in an attempt to identify target genes responsible for apoptosis induced by etoposide, a p53 activating topoisomerase II inhibitor used clinically as an antitumor agent. 62 Individual mRNAs whose mass changed significantly were identified after screening oligonucleotide arrays capable of detecting 6591 unique human mRNA species. 12 (Nine induced and three repressed) of the etoposide-responsive genes were further studied by Northern analysis and an agreement rate of 92%, was reached. Among the 12 genes studied, two (WAF1/p21 and PCNA) are known p53 regulatory genes, two (glutathione peroxidase and S100A2 calcium-binding protein) appear to be the novel p53 target genes and the others appear to be p53-independent. Based upon these findings, the signalling pathways that possibly mediate etoposide-induced apoptosis are proposed.
Insights
This study used DNA chip technology to find genes involved in etoposide-induced apoptosis. Researchers identified novel p53 target genes and proposed signaling pathways mediating this cell death process.
Area of Science:
- Molecular Biology
- Genomics
- Cancer Research
Background:
- Etoposide is a clinical antitumor agent that inhibits topoisomerase II and activates p53.
- Understanding genes regulated by p53 is crucial for cancer therapy.
- DNA chip technology offers a high-throughput method for gene expression analysis.
Purpose of the Study:
- To identify genes targeted by p53 in response to etoposide treatment using DNA chip technology.
- To distinguish between p53-dependent and p53-independent genes affected by etoposide.
- To propose signaling pathways involved in etoposide-induced apoptosis.
Main Methods:
- Oligonucleotide arrays were used to screen 6591 human mRNA species.
- Differential gene expression analysis identified mRNAs with significant mass changes.
- Northern blot analysis was performed on selected candidate genes for validation.
Main Results:
- DNA chip analysis identified 62 differentially expressed mRNAs in response to etoposide.
- Northern analysis confirmed 12 etoposide-responsive genes with 92% agreement.
- Two known p53 regulatory genes (WAF1/p21, PCNA) and two potential novel p53 target genes (glutathione peroxidase, S100A2) were identified.
Conclusions:
- Etoposide treatment affects a specific set of genes, including known and novel p53 targets.
- The study proposes signaling pathways mediating etoposide-induced apoptosis.
- Findings contribute to understanding p53-mediated cellular responses to chemotherapy.