Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1994 Feb;93(2):759–767. doi: 10.1172/JCI117030

Expression of inducible stress protein 70 in rat heart myogenic cells confers protection against simulated ischemia-induced injury.

R Mestril 1, S H Chi 1, M R Sayen 1, K O'Reilly 1, W H Dillmann 1
PMCID: PMC293923  PMID: 8113409

Abstract

Myocardial ischemia markedly increases the expression of several members of the stress/heat shock protein (HSP) family, especially the inducible HSP70 isoforms. Increased expression of HSP70 has been shown to exert a protective effect against a lethal heat shock. We have examined the possibility of using this resistance to a lethal heat shock as a protective effect against an ischemic-like stress in vitro using a rat embryonic heart-derived cell line H9c2 (2-1). Myogenic cells in which the heat shock proteins have been induced by a previous heat shock are found to become resistant to a subsequent simulated ischemic stress. In addition, to address the question of how much does the presence of the HSP70 contribute to this protective effect, we have generated stably transfected cell lines overexpressing the human-inducible HSP70. Embryonal rat heart-derived H9c2(2-1) cells were used for this purpose. This stably transfected cell line was found to be significantly more resistant to an ischemic-like stress than control myogenic cells only expressing the selectable marker (neomycin) or the parental cell line H9c2(2-1). This finding implicates the inducible HSP70 protein as playing a major role in protecting cardiac cells against ischemic injury.

Full text

PDF
759

Images in this article

761Image
on p.761
762Image
on p.762
762Image
on p.762
762Image
on p.762
763Image
on p.763
763Image
on p.763
763Image
on p.763
764Image
on p.764

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abravaya K., Myers M. P., Murphy S. P., Morimoto R. I. The human heat shock protein hsp70 interacts with HSF, the transcription factor that regulates heat shock gene expression. Genes Dev. 1992 Jul;6(7):1153–1164. doi: 10.1101/gad.6.7.1153. [DOI] [PubMed] [Google Scholar]
  2. Angelidis C. E., Lazaridis I., Pagoulatos G. N. Constitutive expression of heat-shock protein 70 in mammalian cells confers thermoresistance. Eur J Biochem. 1991 Jul 1;199(1):35–39. doi: 10.1111/j.1432-1033.1991.tb16088.x. [DOI] [PubMed] [Google Scholar]
  3. Baler R., Welch W. J., Voellmy R. Heat shock gene regulation by nascent polypeptides and denatured proteins: hsp70 as a potential autoregulatory factor. J Cell Biol. 1992 Jun;117(6):1151–1159. doi: 10.1083/jcb.117.6.1151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chirico W. J., Waters M. G., Blobel G. 70K heat shock related proteins stimulate protein translocation into microsomes. Nature. 1988 Apr 28;332(6167):805–810. doi: 10.1038/332805a0. [DOI] [PubMed] [Google Scholar]
  6. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  7. Collier N. C., Schlesinger M. J. The dynamic state of heat shock proteins in chicken embryo fibroblasts. J Cell Biol. 1986 Oct;103(4):1495–1507. doi: 10.1083/jcb.103.4.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Currie R. W. Effects of ischemia and perfusion temperature on the synthesis of stress-induced (heat shock) proteins in isolated and perfused rat hearts. J Mol Cell Cardiol. 1987 Aug;19(8):795–808. doi: 10.1016/s0022-2828(87)80390-5. [DOI] [PubMed] [Google Scholar]
  9. Currie R. W., Karmazyn M., Kloc M., Mailer K. Heat-shock response is associated with enhanced postischemic ventricular recovery. Circ Res. 1988 Sep;63(3):543–549. doi: 10.1161/01.res.63.3.543. [DOI] [PubMed] [Google Scholar]
  10. Currie R. W., Tanguay R. M., Kingma J. G., Jr Heat-shock response and limitation of tissue necrosis during occlusion/reperfusion in rabbit hearts. Circulation. 1993 Mar;87(3):963–971. doi: 10.1161/01.cir.87.3.963. [DOI] [PubMed] [Google Scholar]
  11. Deshaies R. J., Koch B. D., Werner-Washburne M., Craig E. A., Schekman R. A subfamily of stress proteins facilitates translocation of secretory and mitochondrial precursor polypeptides. Nature. 1988 Apr 28;332(6167):800–805. doi: 10.1038/332800a0. [DOI] [PubMed] [Google Scholar]
  12. Dillmann W. H., Mehta H. B., Barrieux A., Guth B. D., Neeley W. E., Ross J., Jr Ischemia of the dog heart induces the appearance of a cardiac mRNA coding for a protein with migration characteristics similar to heat-shock/stress protein 71. Circ Res. 1986 Jul;59(1):110–114. doi: 10.1161/01.res.59.1.110. [DOI] [PubMed] [Google Scholar]
  13. Donnelly T. J., Sievers R. E., Vissern F. L., Welch W. J., Wolfe C. L. Heat shock protein induction in rat hearts. A role for improved myocardial salvage after ischemia and reperfusion? Circulation. 1992 Feb;85(2):769–778. doi: 10.1161/01.cir.85.2.769. [DOI] [PubMed] [Google Scholar]
  14. Feder J. H., Rossi J. M., Solomon J., Solomon N., Lindquist S. The consequences of expressing hsp70 in Drosophila cells at normal temperatures. Genes Dev. 1992 Aug;6(8):1402–1413. doi: 10.1101/gad.6.8.1402. [DOI] [PubMed] [Google Scholar]
  15. Hickey E., Brandon S. E., Sadis S., Smale G., Weber L. A. Molecular cloning of sequences encoding the human heat-shock proteins and their expression during hyperthermia. Gene. 1986;43(1-2):147–154. doi: 10.1016/0378-1119(86)90018-1. [DOI] [PubMed] [Google Scholar]
  16. Hunt C., Morimoto R. I. Conserved features of eukaryotic hsp70 genes revealed by comparison with the nucleotide sequence of human hsp70. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6455–6459. doi: 10.1073/pnas.82.19.6455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Iwaki K., Chi S. H., Dillmann W. H., Mestril R. Induction of HSP70 in cultured rat neonatal cardiomyocytes by hypoxia and metabolic stress. Circulation. 1993 Jun;87(6):2023–2032. doi: 10.1161/01.cir.87.6.2023. [DOI] [PubMed] [Google Scholar]
  18. Jennings R. B., Hawkins H. K., Lowe J. E., Hill M. L., Klotman S., Reimer K. A. Relation between high energy phosphate and lethal injury in myocardial ischemia in the dog. Am J Pathol. 1978 Jul;92(1):187–214. [PMC free article] [PubMed] [Google Scholar]
  19. Jennings R. B., Reimer K. A., Steenbergen C. Myocardial ischemia revisited. The osmolar load, membrane damage, and reperfusion. J Mol Cell Cardiol. 1986 Aug;18(8):769–780. doi: 10.1016/s0022-2828(86)80952-x. [DOI] [PubMed] [Google Scholar]
  20. Johnston R. N., Kucey B. L. Competitive inhibition of hsp70 gene expression causes thermosensitivity. Science. 1988 Dec 16;242(4885):1551–1554. doi: 10.1126/science.3201244. [DOI] [PubMed] [Google Scholar]
  21. Lemasters J. J., DiGuiseppi J., Nieminen A. L., Herman B. Blebbing, free Ca2+ and mitochondrial membrane potential preceding cell death in hepatocytes. Nature. 1987 Jan 1;325(6099):78–81. doi: 10.1038/325078a0. [DOI] [PubMed] [Google Scholar]
  22. Li G. C., Li L. G., Liu Y. K., Mak J. Y., Chen L. L., Lee W. M. Thermal response of rat fibroblasts stably transfected with the human 70-kDa heat shock protein-encoding gene. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1681–1685. doi: 10.1073/pnas.88.5.1681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mehta H. B., Popovich B. K., Dillmann W. H. Ischemia induces changes in the level of mRNAs coding for stress protein 71 and creatine kinase M. Circ Res. 1988 Sep;63(3):512–517. doi: 10.1161/01.res.63.3.512. [DOI] [PubMed] [Google Scholar]
  24. Riabowol K. T., Mizzen L. A., Welch W. J. Heat shock is lethal to fibroblasts microinjected with antibodies against hsp70. Science. 1988 Oct 21;242(4877):433–436. doi: 10.1126/science.3175665. [DOI] [PubMed] [Google Scholar]
  25. Seip W. F., Evans G. L. Atmospheric analysis and redox potentials of culture media in the GasPak System. J Clin Microbiol. 1980 Mar;11(3):226–233. doi: 10.1128/jcm.11.3.226-233.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Summerhayes I. C., Lampidis T. J., Bernal S. D., Nadakavukaren J. J., Nadakavukaren K. K., Shepherd E. L., Chen L. B. Unusual retention of rhodamine 123 by mitochondria in muscle and carcinoma cells. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5292–5296. doi: 10.1073/pnas.79.17.5292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Velazquez J. M., Lindquist S. hsp70: nuclear concentration during environmental stress and cytoplasmic storage during recovery. Cell. 1984 Mar;36(3):655–662. doi: 10.1016/0092-8674(84)90345-3. [DOI] [PubMed] [Google Scholar]
  28. Welch W. J., Feramisco J. R. Rapid purification of mammalian 70,000-dalton stress proteins: affinity of the proteins for nucleotides. Mol Cell Biol. 1985 Jun;5(6):1229–1237. doi: 10.1128/mcb.5.6.1229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Yellon D. M., Pasini E., Cargnoni A., Marber M. S., Latchman D. S., Ferrari R. The protective role of heat stress in the ischaemic and reperfused rabbit myocardium. J Mol Cell Cardiol. 1992 Aug;24(8):895–907. doi: 10.1016/0022-2828(92)91102-b. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES