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Docetaxel induced gene expression patterns in head and neck squamous cell carcinoma using cDNA microarray and PowerBlot.
- Source :
-
Clinical cancer research : an official journal of the American Association for Cancer Research [Clin Cancer Res] 2002 Dec; Vol. 8 (12), pp. 3910-21. - Publication Year :
- 2002
-
Abstract
- Purpose: The purpose is to identify gene expression patterns induced by docetaxelin head and neck squamous carcinoma (HNSCC) cells using high throughput techniques.<br />Experimental Design: HNSCC cells were treated with docetaxel or solvent. After mRNA extraction, cDNA fluorescent (Cy3 or Cy5)-labeled probes were synthesized. Then, Cy3 and Cy5-labeled samples were hybridized onto a microarray slide. The fluorescent images were scanned and analyzed for quantification. PowerBlot immunoblotting technique was used to measure protein expression level. Using this dual approach, we focused on genes in established pathways (cell cycle, apoptosis, angiogenesis, and signal transduction) of tumorigenesis and confirmed these results with conventional techniques.<br />Results: Using cDNA microarray, we found that docetaxel altered the expression of >100 genes in HNSCC cells. A total of 153 of 1191 genes was found to have altered expression in either HN12 (n = 102), HN30 (n = 72), or both (n = 21) by docetaxel. For the PowerBlot analysis, a subset of genes (n = 46) in the cDNA microarray analysis and an additional 98 genes in the cell cycle, apoptosis, angiogenesis, and signal transduction pathways were chosen. We found that PowerBlot data agreed with cDNA microarray in 65% of genes examined. The expression of a cell cycle inhibitor (p19) and promoters (cyclin A, cyclin B1, and cyclin E2F) were increased and decreased, respectively. Apoptosis induced by docetaxel was independent of p53 and, in part, related to increased Fas expression. Both vascular endothelial growth factor secretion and basic fibroblast growth factor expression were inhibited by docetaxel, whereas thrombospondin-1 expression was increased by docetaxel. Epidermal growth factor receptor, activated epidermal growth factor receptor, and activated c-Jun NH(2)-terminal kinase expression was lowered by docetaxel. Activated extracellular signal-regulated kinase was elevated by docetaxel, but not total extracellular signal-regulated kinase levels.<br />Conclusions: The identification of altered gene expression induced by docetaxel demonstrates additional biological activity in HNSCC cells, and the altered expression of these genes may serve as potential biomarkers to both predict clinical activity and provide information regarding potential efficacy of adding novel agents.
- Subjects :
- Annexin A5 metabolism
Apoptosis drug effects
Blotting, Western
Carcinoma, Squamous Cell metabolism
DNA Primers chemistry
Docetaxel
Endothelial Growth Factors metabolism
Head and Neck Neoplasms metabolism
Humans
Intercellular Signaling Peptides and Proteins metabolism
Lymphokines metabolism
Neoplasm Proteins metabolism
Oligonucleotide Array Sequence Analysis
RNA, Neoplasm metabolism
Reverse Transcriptase Polymerase Chain Reaction
Vascular Endothelial Growth Factor A
Vascular Endothelial Growth Factors
fas Receptor metabolism
Antineoplastic Agents, Phytogenic pharmacology
Carcinoma, Squamous Cell genetics
Gene Expression Profiling
Gene Expression Regulation, Neoplastic drug effects
Head and Neck Neoplasms genetics
Neoplasm Proteins genetics
Paclitaxel analogs & derivatives
Paclitaxel pharmacology
Taxoids
Subjects
Details
- Language :
- English
- ISSN :
- 1078-0432
- Volume :
- 8
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Clinical cancer research : an official journal of the American Association for Cancer Research
- Publication Type :
- Academic Journal
- Accession number :
- 12473607