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KMID : 0857020060210010338
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Kosin Medical Journal
2006 Volume.21 No. 1 p.338 ~ p.343
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Analysis of gene expression profile in 10-Hydroxycamptothecin-treated human leukemia HL-60 cells using oligonucleotide microarray
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Jeong In-Cheol
Cho Moo-Youn
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Abstract
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Background : Camptothecin is an antitumor alkaloid that has been isolated from the Chinese tree, Camptotheca acuminata.. DNA topoisomerase I has been firmly established as the molecular target of the camptothecin group of anticancer drugs. These drugs include 10-hydroxycamptothecin(10-CPT), topotecan, 9-aminocamptothecin, and irinotecan. Many of them are now in clinical trials and are showing activity against a wide variety of solid human malignancies. DNA chip microarray technology can be used to establish associations between characteristic gene expression patterns and molecular responses to drug therapy. This study was conducted to determine whether pretreatment of HL-60 human leukemia cells with the topoisomerase I - directed drug 10-CPT regulates gene expression and to investigate the activity of topoisomerase mediated by 10-CPT in nuclear extract from HL-60 human leukemia cells.
Methods :We have conducted experiments on cell cytotoxicity in drug-treated cellls, topoisomerase purification, enzyme assay using agarose gel electrophoresis and oligo chip microarray analysis, respectively.
Results : Treatment of HL-60 cells with 10-CPT resulted in marked inhibition of type I topoisomerase activity. 10-CPT IC(50) values was 8 nM for HL-60 cells. Thirty-one genes that were up-regulated(18 genes) and down-regulated(13 genes) in 10-CPT treated HL-60 cells were found.
Conclusion :Our results suggest that the various expression profiles of newly identified genes in 10-CPT treated human leukemia cells by oligonucleotide microarray can be potentially used as novel therapeutic markers.
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KEYWORD
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10-hydroxycamptothecin(10-CPT), Topoisomerase, Expression profile, Oligonucleotide microarray
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