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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Notes
The figure thus does not include DNAJA3, whose largest significant change in expression was a decrease in expression of 0.75-fold relative to control at the T = 3 time point.
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The views, opinions, and findings contained in this publication are those of the authors and should not be construed as an official USUHS or US Department of the Army position, policy, or decision, unless so designated by other documentation.
Approved for public release; distribution unlimited.
Funding was provided by the US Army Medical Research and Materiel Command (USAMRMC). National Heart, Lung, and Blood Institute Grants HL-64104 and HL-072114 also supported this work.
The authors thank Dr. Michael Sawka of the US Army Research Institute of Environmental Medicine, Natick, MA, USA and Dr. Karen Fairchild of the University of Virginia, Charlottesville, VA, USA for critical review of this manuscript. The authors would also like to thank Dr. Jaques Reifman, whose input contributed to the development of the software used to analyze these data.
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Sonna, L.A., Kuhlmeier, M.M., Khatri, P. et al. A microarray analysis of the effects of moderate hypothermia and rewarming on gene expression by human hepatocytes (HepG2). Cell Stress and Chaperones 15, 687–702 (2010). https://doi.org/10.1007/s12192-010-0181-2
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DOI: https://doi.org/10.1007/s12192-010-0181-2