Abstract
The upregulation of stress proteins is an important step in the cellular defense against various endogenous and exogenous stressors, and the stress response systems are therefore essential to cell maintenance and survival. In order to evaluate the cellular capability to cope with exogenous stressors, we investigated stress response profiles in cultured human fibroblasts exposed to short-term heat shock or oxidative stress. Quantitative PCR analysis as well as Western blotting demonstrated a specific upregulation of Hsp70 mRNA and protein after heat shock, while heme oxygenase 1 (HO1 or Hsp32) mRNA and protein was specifically upregulated following H2O2-induced oxidative stress. Hsp70 and HO1 should therefore be considered as useful biomarkers for acute heat stress and oxidative stress, respectively. Moreover, the results indicate that the expression of Hsp70 and HO1 in response to exogenous stressors may be regulated through different mechanisms.
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References
Richter-Landsberg C, Goldbaum O (2003) Stress proteins in neural cells: functional roles in health and disease. Cell Mol Life Sci 60:337–349
Unno K, Asakura H, Shibuya Y, Kaihou M, Fukatsu H, Okada S, Oku N (2002) Stress response caused by chronic alcohol intake in aged rat brain. Alcohol Clin Exp Res 26:1017–1023
Verbeke P, Fonager J, Clark BF, Rattan SI (2001) Heat shock response and ageing: mechanisms and applications. Cell Biol Int 25:845–857
Westerheide SD, Morimoto RI (2005) Heat shock response modulators as therapeutic tools for diseases of protein conformation. J Biol Chem 280:33097–33100
Calabrese V, Lodi R, Tonon C, D’Agata V, Sapienza M, Scapagnini G, Mangiameli A, Pennisi G, Stella AM, Butterfield DA (2005) Oxidative stress, mitochondrial dysfunction and cellular stress response in Friedreich’s ataxia. J Neurol Sci 233:145–162
Sonna LA, Fujita J, Gaffin SL, Lilly CM (2002) Invited review: Effects of heat and cold stress on mammalian gene expression. J Appl Physiol 92:1725–1742
Fourie AM, Peterson PA, Yang Y (2001) Characterization and regulation of the major histocompatibility complex-encoded proteins Hsp70-Hom and Hsp70-1/2. Cell Stress Chaperones 6:282–295
Gabai VL, Meriin AB, Yaglom JA, Volloch VZ, Sherman MY (1998) Role of Hsp70 in regulation of stress-kinase JNK: implications in apoptosis and aging. FEBS Lett 438:1–4
Stege GJ, Li GC, Li L, Kampinga HH, Konings AW (1994) On the role of hsp72 in heat-induced intranuclear protein aggregation. Int J Hyperthermia 10:659–674
Bruening W, Roy J, Giasson B, Figlewicz DA, Mushynski WE, Durham HD (1999) Up-regulation of protein chaperones preserves viability of cells expressing toxic Cu/Zn-superoxide dismutase mutants associated with amyotrophic lateral sclerosis. J Neurochem 72:693–699
Elbirt KK, Bonkovsky HL (1999) Heme oxygenase: recent advances in understanding its regulation and role. Proc Assoc Am Physicians 111:438–447
Choi AM, Alam J (1996) Heme oxygenase-1: function, regulation, and implication of a novel stress-inducible protein in oxidant-induced lung injury. Am J Respir Cell Mol Biol 15:9–19
Stocker R (1990) Induction of haem oxygenase as a defence against oxidative stress. Free Radic Res Commun 9:101–112
Clark JE, Foresti R, Green CJ, Motterlini R (2000) Dynamics of haem oxygenase-1 expression and bilirubin production in cellular protection against oxidative stress. Biochem J 348(Pt 3):615–619
Poss KD, Tonegawa S (1997) Reduced stress defense in heme oxygenase 1-deficient cells. Proc Natl Acad Sci U S A 94:10925–10930
Scapagnini G, D’Agata V, Calabrese V, Pascale A, Colombrita C, Alkon D, Cavallaro S (2002) Gene expression profiles of heme oxygenase isoforms in the rat brain. Brain Res 954:51–59
Heid CA, Stevens J, Livak KJ, Williams PM (1996) Real time quantitative PCR. Genome Res 6:986–994
Winer J, Jung CK, Shackel I, Williams PM (1999) Development and validation of real-time quantitative reverse transcriptase-polymerase chain reaction for monitoring gene expression in cardiac myocytes in vitro. Anal Biochem 270:41–49
Acknowledgements
The authors would like to thank lector Thomas J. Corydon at the Institute for Human Genetics, University of Aarhus, for advice, support and use of cell culturing facilities. This work has been supported by The Danish Medical Research Council; Aarhus University Hospital’s Research Initiative; Institute of experimental Clinical Research, University of Aarhus; Faculty of Health Sciences, University of Aarhus.
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Pedersen, C.B., Gregersen, N. (2010). Stress Response Profiles in Human Fibroblasts Exposed to Heat Shock or Oxidative Stress. In: Bross, P., Gregersen, N. (eds) Protein Misfolding and Cellular Stress in Disease and Aging. Methods in Molecular Biology, vol 648. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-756-3_10
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DOI: https://doi.org/10.1007/978-1-60761-756-3_10
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