A microarray analysis of the effects of moderate hypothermia and rewarming on gene expression by human hepatocytes (HepG2) |
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Authors: | Larry A Sonna Matthew M Kuhlmeier Purvesh Khatri Dechang Chen and Craig M Lilly |
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Institution: | (1) Division of Pulmonary and Critical Care Medicine, University of Maryland School of Medicine, Baltimore, MD 21201, USA;(2) Thermal and Mountain Medicine Division, US Army Research Institute of Environmental Medicine, Natick, MA 01760, USA;(3) Department of Computer Science, Wayne State University, Detroit, MI 48202, USA;(4) Division of Epidemiology and Biostatistics, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20814, USA;(5) Division of Allergy, Pulmonary, and Critical Care Medicine, Department of Medicine, University of Massachusetts School of Medicine, Worcester, MA 01655, USA |
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Abstract: | The gene expression changes produced by moderate hypothermia are not fully known, but appear to differ in important ways from
those produced by heat shock. We examined the gene expression changes produced by moderate hypothermia and tested the hypothesis
that rewarming after hypothermia approximates a heat-shock response. Six sets of human HepG2 hepatocytes were subjected to
moderate hypothermia (31°C for 16 h), a conventional in vitro heat shock (43°C for 30 min) or control conditions (37°C), then
harvested immediately or allowed to recover for 3 h at 37°C. Expression analysis was performed with Affymetrix U133A gene
chips, using analysis of variance-based techniques. Moderate hypothermia led to distinct time-dependent expression changes,
as did heat shock. Hypothermia initially caused statistically significant, greater than or equal to twofold changes in expression
(relative to controls) of 409 sequences (143 increased and 266 decreased), whereas heat shock affected 71 (35 increased and
36 decreased). After 3 h of recovery, 192 sequences (83 increased, 109 decreased) were affected by hypothermia and 231 (146
increased, 85 decreased) by heat shock. Expression of many heat shock proteins was decreased by hypothermia but significantly
increased after rewarming. A comparison of sequences affected by thermal stress without regard to the magnitude of change
revealed that the overlap between heat and cold stress was greater after 3 h of recovery than immediately following thermal
stress. Thus, while some overlap occurs (particularly after rewarming), moderate hypothermia produces extensive, time-dependent
gene expression changes in HepG2 cells that differ in important ways from those induced by heat shock. |
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Keywords: | Cold stress Cell stress response DNA microarray Heat shock proteins |
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