Diverse gene expression pattern during 5-fluorouridine-induced apoptosis.

Abstract

The purpose of this study was to establish experimental conditions to produce apoptosis by the fluorinated pyrimidine 5-fluorouridine and to examine the changes in gene expression that occurred during cell death. HCT-116 colorectal carcinoma cells were exposed to 10 microM 5-fluorouridine alone or in the presence of 1 mM uridine, 30 microM thymidine or both uridine and thymidine. A time-dependent increase in the percentage of apoptotic cells and a decrease in the percentage of viable cells were observed when the cells were treated with 5-fluorouridine in the absence of uridine (p < 0.001) but not in the presence of uridine. cDNA microarray analysis was used to study the expression of 1,200 different genes during apoptosis by 5-flurouridine. The expression of 33 genes was upregulated by 5-fold or greater at 16 and 24 h of 5-fluorouridine exposure. The largest cluster of upregulated genes included a group of genes classified as growth factors, cytokines and chemokines (e.g. interleukin-3, interleukin-4, B-cell growth factor 1 and stem cell growth factor). The expression of MIC-1 increased up to 100-fold during 5-flurouridine exposure. One hundred and twenty-four genes were downregulated by 5-fold or greater following exposure to 5-fluorouridine. The downregulated genes were distributed throughout the six different classifications on the array. Our data demonstrate a diverse pattern of gene expression during the fluorouridine-induced apoptosis and suggest that mechanisms besides a global inhibition of RNA synthesis/ processing contribute to the RNA-directed cytotoxicity of fluoropyrimidines.

• Authors: Thomas D Schmittgen, Kari A Gissel, Brian A Zakrajsek, B Paige Lawrence, Qian Liu, Eldon R Jupe, Megan R Lerner, Son V Do, Daniel J Brackett
• Journal: International journal of oncology (Int J Oncol) Vol. 27 Issue 2 Pg. 297-306 (Aug 2005) ISSN: 1019-6439 [Print] Greece
• PMID: 16010409 (Publication Type: Comparative Study, Journal Article, Research Support, N.I.H., Extramural, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• RNA, Neoplasm
• 5-fluorouridine
• Thymidine
• Uridine

Topics

• Analysis of Variance
• Apoptosis (drug effects, genetics)
• Cell Survival (drug effects)
• Cluster Analysis
• Colorectal Neoplasms (genetics, pathology, ultrastructure)
• Flow Cytometry
• Gene Expression Profiling
• Gene Expression Regulation, Neoplastic (drug effects, genetics)
• HCT116 Cells
• Humans
• Microscopy, Electron
• Necrosis
• Oligonucleotide Array Sequence Analysis (methods)
• RNA, Neoplasm (genetics, metabolism)
• Reproducibility of Results
• Reverse Transcriptase Polymerase Chain Reaction (methods)
• Thymidine (pharmacology)
• Time Factors
• Up-Regulation (drug effects, genetics)
• Uridine (analogs & derivatives, metabolism, pharmacology)