Exp Mol Med.  2002 Jul;34(3):184-193.

Genome-wide expression profiling of 8-chloroadenosine- and 8-chloro-cAMP-treated human neuroblastoma cells using radioactive human cDNA microarray.

• Park GH
• Choe JG
• Choo HJ
• Park YG
• Sohn JW
• Kim MK

Abstract

Previous reports raised question as to whether 8-chloro-cyclic adenosine 3,5-monophosphate (8-Cl-cAMP) is a prodrug for its metabolite, 8-Cl-adenosine which exerts growth inhibition in a broad spectrum of cancer cells. The present study was carried out to clarify overall cellular affects of 8-Cl-cAMP and 8-Cl-adenosine on SK-N-DZ human neuroblastoma cells by ystematically characterizing gene expression using radioactive human cDNA microarray. Microarray was prepared with PCR-amplified cDNA of 2,304 known genes spotted on nylon membranes, employing (1)P-labeled cDNAs of SK-N-DZ cells as a probe. the expression levels of approximately 100 cDNAs, representing about 8% of the total DNA elements on the array, were altered in 8-Cl-adenosine- or 8-Cl-cAMP-treated cells, respectively. The genome-wide expression of the two samples exhibited partial overlaps; different sets of up-regulated genes but the same set of down-regulated genes. 8-Cl-adenosine treatment up- egulated genes involved in differentiation and development (LIM protein, connexin 26, neogenin, neurofilament triplet L protein and p21( WAF1/CIP1)) and immune response such as natural killer cells protein 4, and down-regulated ones involved in proliferation and transformation (transforming growth factor-beta, DYRK2, urokinase-type plasminogen activator and proteins involved in transcription and translation) which were in close parallel with those by 8-Cl-cAMP. Our results indicated that the two drugs shared common genomic pathways for the down-regulation of certain genes, but used distinct pathways for the up-regulation of different gene clusters. Based on the findings, we suggest that the anti-cancer activity of 8-Cl-cAMP results at least in part through 8-Cl-adenosine. Thus, the systematic use of DNA arrays can provide insight into the dynamic cellular pathways involved in anticancer activities of chemotherapeutics.

Keyword

8-Cl-adenosine; 8-Cl-cAMP; anticancer activity; radioactive cDNA microarray; genome-wide expression

MeSH Terms

2-Chloroadenosine/*analogs & derivatives/chemistry/*pharmacology
8-Bromo Cyclic Adenosine Monophosphate/*analogs & derivatives/chemistry/*pharmacology
Antineoplastic Agents/chemistry/*pharmacology
Blotting, Western
*Gene Expression Profiling
Gene Expression Regulation, Neoplastic/*drug effects
Genome, Human
Human
Neuroblastoma/*genetics
Oligonucleotide Array Sequence Analysis
Reproducibility of Results
Tumor Cells, Cultured
Up-Regulation/drug effects