Abstract
The effects of the oxidizing agent t-butylhydroperoxide (t-BHP) were investigated on three human cell lines of different origin and growth features (A431 epithelial cells, ADF astrocytoma cells and U937 leukemic cells) using electron microscopy and electron paramagnetic resonance spectroscopy. The results indicate that important biophysical and ultrastructural modifications are induced in the plasma and mitochondrial membranes of these cells and that these changes can ultimately lead to cell death. In addition, the cell cytoskeleton also appears to be a target of hydroperoxide-mediated stress. In particular, all three cell types undergo cytoskeletal alterations leading to surface blebbing, a typical characteristic of cell damage. However, the timing and extent of this damage as well as that occurring at the mitochondrial and plasma membrane levels seems to be different: cells with weak (ADF) or absent (U937) cell-to-cell and cell-substrate contacts and a poorly developed cytoskeleton appear to be more susceptible than other cell types (e.g., A431) to t-BHP-mediated injury. These diverse cell susceptibilities to hydroperoxide-mediated oxidative stress could thus depend upon cell histotype-associated growth featurs.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- t-BHP:
-
tert-butylhydroperoxide
- DMEM:
-
Dulbecco's Modified Eagle's Medium
- EPR:
-
electron paramagnetic resonance
- GSH:
-
reduced glutathione
- 5-NSA:
-
nitroxystearic acid
- PBS:
-
phosphate-buffered saline
References
Bellomo G, Mirabelli F. Oxidative stress and cytoskeletal alterations. Ann NY Acad Sci. 1992;663:97–109.
Bellomo G, Jewell SA, Thor H, Orrenius S. Regulation of intracellular calcium compartmentation: studies with isolated hepatocytes and t-butyl hydroperoxide. Proc Natl Acad Sci USA. 1982a;79:6842–6.
Bellomo G, Jewell SA, Orrenius S. The metabolism of menadione impairs the ability of rat liver mitochondria to take up and retain calcium. J Biol Chem. 1982b;257:11558–62.
Coakley WT. Hyperthermia effects on the cytoskeleton and on cell morphology. In: Bowler K, Fuller BJ, eds. Temperature and animal cells. Cambridge: Company of Biologists Ltd; 1987:187–211.
Duncan CJ, Shamsadeen N. Ultrastructural changes in mitochondria during rapid damage triggered by calcium. In: Duncan CJ, ed. Calcium, oxygen radicals and cellular damage. Cambridge: Cambridge University Press; 1993:149–64.
Hynes RO. Integrins: versatility, modulation, and signalling in cell adhesion. Cell. 1992;69:11–25.
Imberti R, Nieminen AL, Herman B, Lemasters JJ. Mitochondrial and glycolytic dysfunction in lethal injury to hepatocytes by t-butylhydroperoxide: protection by fructose, cyclosporin A and trifluoperazine. J Pharmacol Exp Ther. 1993;265:392–400.
Gutteridge JMC. Free radicals in disease processes: a complilation of cause and consequence. Free Rad Res Commun. 1993;19:141–58.
Jewell SA, Bellomo G, Thor H, Orrenius S, Smith MT. Bleb formation in hepatocytes during drug metabolism is associated with alterations in intracellular thiol and calcium ion homeotasis. Science. 1982;217:1257–9.
Kennedy CH, Church DF, Winston GW, Pryor WA. tert-Butyl hyperoxide-induced radical production in rat liver mitochondria. Free Rad Biol Med. 1992;12:381–7.
Lehningher AL, Vercesi AE, Bababunmi EA. Regulation of Ca2+ release from mitochondria by the oxidation reduction state of pyridine nucleotides. Proc Natl Acad Sci USA. 1978;75:1690–4.
Lotscher HR, Winterhalter KH, Carafoli E, Richter C. Hydroperoxides can modulate the redox state of pyridine nucleotides and calcium balance in rat liver mitochondria. Proc Natl Acad Sci USA. 1979;76:4340–4.
Luna EJ, Hitt AL. Cytoskeleton-plasma membrane interactions. Science. 1992;258:955–64.
Malorni W, Iosi F, Marabelli F, Bellomo G. Cytoskeleton as target in menadione-induced oxidative stress in cultured mammalian cells: alterations underlying surface bleb formation. Chem Biol Interact. 1991;80:217–36.
Malorni W, Iosi F, Donelli G, Caprari P, Salvati AM, Cianciulli P. A new striking morphologic feature for the human erythrocyte in hereditary spherocytosis: the bledding pattern. Blood. 1993;81:2821–2.
McConnell HM, Gaffney McFarland B. Cytochalasin B disrupts the association of filamentous web and plasma membrane in hepatocytes. Q Rev Biophys. 1970;3:91–136.
Orrenius S, McConkey DJ, Bellomo G, Nicotera P. Role of calcium in toxic cell killing. Trends Pharm Sci. 1989;10:281–5.
Santini MT, Indovina PL, Hausman RE. Changes in myoblast membrane order during differentiation as measured by EPR. Biochim Biophys Acta. 1987;896:19–25.
Saville B. A scheme for the colorimetric determination of microgram amounts of thiols. Analyst. 1958;83:670–2.
Sies H. Oxidative stress: introductory remarks. In: Sies H, ed. Oxidative stress. London: Academic Press; 1985:1–8.
Sies H. Oxidative stress: from basic research to clinical application. Am J Med. 1991;91(3C):31–8.
Smith MT, Thor H, Jewell SA, Bellomo G, Sandy MS, Orrenius S. Free radical-induced changes in the surface morphology of isolated hepatocytes. In: Armstrong D, et al. Free radicals in molecular biology, aging and disease. New York: Raven Press; 1984:103–18.
Van Den Berg JJM, Op den Kamp JAF, Bertram HL, Roelofsen B, Kuypers FA. Kinetics and site specificity of hydroperoxide-induced oxidative damage in red blood cells. Free Rad Biol Med. 1992;12:487–98.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Malorni, W., Rainaldi, G., Rivabene, R. et al. Different susceptibilities to cell death induced by t-butylhydroperoxide could depend upon cell histotype-associated growth features. Cell Biol Toxicol 10, 207–218 (1994). https://doi.org/10.1007/BF00756761
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00756761