Abstract
Dysregulation of epigenetic control is an important participant in carcinogenesis. The PML/RAR α translocation in acute promyelocytic leukemia (APL) is an example where the resultant fusion protein recruits histone deacetylase complexes to target genes resulting in their inappropriate transcriptional repression. All-trans-retinoic acid (ATRA) acts as a ligand that relieves this repression and produces an epigenetic transcriptional reprogramming of the cancer cell. CpG island microarrays were used to analyze the DNA methylation and histone acetylation state of the human APL cell line NB4 before and after differentiation with ATRA as well as normal peripheral blood mononuclear cells (PBMC). Over 70 CpG islands within 1 kb of transcription start of a known gene are aberrantly methylated in NB4 cells compared with PBMC; however, no changes in cytosine methylation were detected following ATRA-induced differentiation. With respect to histone H4 acetylation, over 100 single-copy CpG islands within 1 kb of transcription start of a known human gene became hyperacetylated following ATRA-induced differentiation. One CpG island was aberrantly methylated in NB4 cells, but became hyperacetylated and was induced following ATRA treatment and was associated with the HoxA1 gene, suggesting it may be a target gene of ATRA in APL. In addition to single-copy sequences, a selective increase in acetylation was detected in satellite DNA when compared with other high-copy sequences, such as Alu or rDNA. In summary, ATRA stimulates complex epigenomic changes during leukemic cell differentiation, and monitoring these changes may help to identify new targets of epigenetic dysfunction.
Footnotes
-
National Institutes of Health Grant A091351 to B.W.F. and the National Institute of Environmental Health Sciences Center Grant ES06694 supported this work. M.M.O. was supported, in part, by a Cancer Biology Training grant from the National Institutes of Health.
-
doi:10.1124/jpet.104.072488.
-
ABBREVIATIONS: RAR, retinoic acid receptor; APL, acute promyelocytic leukemia; ATRA, all-trans-retinoic acid; PBMC, peripheral blood mononuclear cell(s); PCR, polymerase chain reaction; SSC, standard saline citrate; Cy3, cyanine 3; Cy5, cyanine 5; ChIP, chromatin immunoprecipitation; IP, input; McrBC, methylation-sensitive restriction enzyme; PIPES, 1,4-piperazinediethanesulfonic acid; TS, transcription start; kb, kilobase(s); kbp, kilobase pair(s).
- Received June 9, 2004.
- Accepted August 6, 2004.
- The American Society for Pharmacology and Experimental Therapeutics
JPET articles become freely available 12 months after publication, and remain freely available for 5 years.Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page.
|