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1 Thermal and Mountain Medicine Division, US Army Research Institute of Environmental Medicine, Natick 01760; and 2 Division of Pulmonary and Critical Care Medicine and 3 Partners Gene Array Technology Center, Brigham and Women's Hospital/Harvard Medical School, Boston, Massachusetts 02115
We studied the effect of heat shock on gene expression by normal human cells. Peripheral blood mononuclear cells (PBMCs) were obtained from healthy adults. Paired samples from each subject were subjected to either 20 min of heat shock (43°C) or control (37°C) conditions and then returned to 37°C. RNA was isolated 160 min later, and five representative samples were analyzed on Affymetrix gene chip arrays containing ~12,600 probes. A biologically meaningful effect was defined as a statistically significant, twofold or greater difference in expression of sequences that were detected in all five experiments under control (downregulated sequences) or heat shock (upregulated sequences) conditions. Changes occurred in 395 sequences (227 increased by heat shock, 168 decreased), representing 353 Unigene numbers, in every functional category previously implicated in the heat shock response. By RT-PCR, we confirmed the findings for one upregulated sequence (Rad, a G protein) and one downregulated sequence (osteopontin, a cytokine). We conclude that heat shock causes extensive gene expression changes in PBMCs, affecting all functional categories of the heat shock response.
apoptosis; gene chip array technology; cell stress response
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