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Gene expression analysis of primary human myeloid precursor cells treated with O6-benzylguanine (6BG) and Temozolomide (TMZ)


ABSTRACT: For the comparison of 6BG/TMZ with control sample, the predominant annotation among the upregulated genes is M-bM-^@M-^XapoptosisM-bM-^@M-^Y. The majority of the downregulated genes is assigned to heat shock proteins and proteins which bind unfolded proteins. To look at early changes in gene expression, primary human myeloid precursor cells (40 x 10^6 per sample) derived from 3 pooled CD34+ products were treated for 18 hours with control (vehicle), 6BG, TMZ, or 6BG/TMZ and cell pellets flash frozen. Total RNA were isolated at Miltenyi Biotec (Cologne, Germany) and bioinformatics analysis of four microarray datasets was performed by their Bioinformatics Group. The direct comparisons were: 6BG/TMZ vs Control, TMZ vs 6BG, 6BG/TMZ vs 6BG, 6BG/TMZ vs TMZ. A two-dye competitive hybridization of mRNAs derived from differently treated human cells in comparison to a reference mRNA derived from cells which underwent a different treatment was conducted. After treatment with two different drugs or a combination of both drugs, respectively, RNA was extracted from the cells and hybridized against the corresponding reference mRNA. As microarray platform, the PIQORM-bM-^DM-" Cell Death Microarray with 494 probes was used.

ORGANISM(S): Homo sapiens

SUBMITTER: Silvia Rueberg 

PROVIDER: E-GEOD-44122 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Temozolomide-mediated DNA methylation in human myeloid precursor cells: differential involvement of intrinsic and extrinsic apoptotic pathways.

Wang Haiyan H   Cai Shanbao S   Ernstberger Aaron A   Bailey Barbara J BJ   Wang Michael Z MZ   Cai Wenjing W   Goebel W Scott WS   Czader Magdalena B MB   Crean Colin C   Suvannasankha Attaya A   Shokolenkoc Inna I   Wilson Glenn L GL   Baluyut Arthur R AR   Mayo Lindsey D LD   Pollok Karen E KE  

Clinical cancer research : an official journal of the American Association for Cancer Research 20130327 10


<h4>Purpose</h4>An understanding of how hematopoietic cells respond to therapy that causes myelosuppression will help develop approaches to prevent this potentially life-threatening toxicity. The goal of this study was to determine how human myeloid precursor cells respond to temozolomide (TMZ)-induced DNA damage.<h4>Experimental design</h4>We developed an ex vivo primary human myeloid precursor cells model system to investigate the involvement of cell-death pathways using a known myelosuppressi  ...[more]

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