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. 1988 Nov;133(2):381–388.

Ultrastructure and dynamics of selective mitochondrial injury in carcinoma cells after doxycycline photosensitization in vitro.

C R Shea 1, D Whitaker 1, G F Murphy 1, T Hasan 1
PMCID: PMC1880784  PMID: 3056023

Abstract

Mechanisms and intracellular sites of photosensitized damage were investigated in cultured MGH-U1 cells treated with doxycycline (DOTC). Cells were examined by phase-contrast, fluorescence, and electron microscopy at various times (15 minutes to 24 hours) after ultraviolet irradiation (320-400 nm). DOTC localized selectively within the mitochondria, as shown by colocalized fluorescence with rhodamine 123 (R123). Photosensitization at 1 J/sq cm caused striking swelling of the mitochondrial matrix and disruption of the cristae, accompanied by loss of the ability of mitochondria to concentrate R123. These changes progressed during the first hour after irradiation, and then were followed by partial recovery of mitochondrial ultrastructure and function. At no time were other organelles seen to be affected. It appears that this selective, photosensitized alteration was a consequence of localized and partially reversible damage to the mitochondrial inner membrane. In contrast, exposure to 2-6 J/sq cm caused irreversible injury and necrosis.

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Selected References

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

  1. Berns M. W., Rattner J. B., Meredith S., Witter M. Current laser microirradiation studies. Ann N Y Acad Sci. 1976 Jan 30;267:160–175. doi: 10.1111/j.1749-6632.1976.tb41606.x. [DOI] [PubMed] [Google Scholar]
  2. Bjellerup M., Ljunggren B. Photohemolytic potency of tetracyclines. J Invest Dermatol. 1985 Apr;84(4):262–264. doi: 10.1111/1523-1747.ep12265336. [DOI] [PubMed] [Google Scholar]
  3. Blank H., Cullen S. I., Catalano P. M. Photosensitivity studies with demethylchlortetracycline and doxycycline. Arch Dermatol. 1968 Jan;97(1):1–2. [PubMed] [Google Scholar]
  4. Chandler D. E., Williams J. A. Intracellular divalent cation release in pancreatic acinar cells during stimulus-secretion coupling. II. Subcellular localization of the fluorescent probe chlorotetracycline. J Cell Biol. 1978 Feb;76(2):386–399. doi: 10.1083/jcb.76.2.386. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Coppola A., Viggiani E., Salzarulo L., Rasile G. Ultrastructural changes in lymphoma cells treated with hematoporphyrin and light. Am J Pathol. 1980 Apr;99(1):175–192. [PMC free article] [PubMed] [Google Scholar]
  6. DU BUY H. G., SHOWACRE J. L. Selective localization of tetracycline in mitochondria of living cells. Science. 1961 Jan 20;133(3447):196–197. doi: 10.1126/science.133.3447.196. [DOI] [PubMed] [Google Scholar]
  7. Garlid K. D., Beavis A. D. Evidence for the existence of an inner membrane anion channel in mitochondria. Biochim Biophys Acta. 1986;853(3-4):187–204. doi: 10.1016/0304-4173(87)90001-2. [DOI] [PubMed] [Google Scholar]
  8. Goldman R. A., Hasan T., Hall C. C., Strycharz W. A., Cooperman B. S. Photoincorporation of tetracycline into Escherichia coli ribosomes. Identification of the major proteins photolabeled by native tetracycline and tetracycline photoproducts and implications for the inhibitory action of tetracycline on protein synthesis. Biochemistry. 1983 Jan 18;22(2):359–368. doi: 10.1021/bi00271a020. [DOI] [PubMed] [Google Scholar]
  9. Hasan T., Khan A. U. Phototoxicity of the tetracyclines: photosensitized emission of singlet delta dioxygen. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4604–4606. doi: 10.1073/pnas.83.13.4604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hasan T., Kochevar I. E., McAuliffe D. J., Cooperman B. S., Abdulah D. Mechanism of tetracycline phototoxicity. J Invest Dermatol. 1984 Sep;83(3):179–183. doi: 10.1111/1523-1747.ep12263531. [DOI] [PubMed] [Google Scholar]
  11. Holland P., Sleamaker K. A cytologic evaluation of intracellular tetracycline fluorescence in normal and neoplastic lymphocytes. Acta Cytol. 1969 Apr;13(4):246–249. [PubMed] [Google Scholar]
  12. Jennings R. B., Schaper J., Hill M. L., Steenbergen C., Jr, Reimer K. A. Effect of reperfusion late in the phase of reversible ischemic injury. Changes in cell volume, electrolytes, metabolites, and ultrastructure. Circ Res. 1985 Feb;56(2):262–278. doi: 10.1161/01.res.56.2.262. [DOI] [PubMed] [Google Scholar]
  13. Johnson L. V., Walsh M. L., Bockus B. J., Chen L. B. Monitoring of relative mitochondrial membrane potential in living cells by fluorescence microscopy. J Cell Biol. 1981 Mar;88(3):526–535. doi: 10.1083/jcb.88.3.526. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jolly S. R., Kane W. J., Bailie M. B., Abrams G. D., Lucchesi B. R. Canine myocardial reperfusion injury. Its reduction by the combined administration of superoxide dismutase and catalase. Circ Res. 1984 Mar;54(3):277–285. doi: 10.1161/01.res.54.3.277. [DOI] [PubMed] [Google Scholar]
  15. Klaunig J. E., Selman S. H., Shulok J. R., Schafer P. J., Britton S. L., Goldblatt P. J. Morphologic studies of bladder tumors treated with hematoporphyrin derivative photochemotherapy. Am J Pathol. 1985 May;119(2):236–243. [PMC free article] [PubMed] [Google Scholar]
  16. Kroon A. M., Dontje B. H., Holtrop M., Van den Bogert C. The mitochondrial genetic system as a target for chemotherapy: tetracyclines as cytostatics. Cancer Lett. 1984 Nov;25(1):33–40. doi: 10.1016/s0304-3835(84)80023-3. [DOI] [PubMed] [Google Scholar]
  17. Kroon A. M., Van den Bogert C. Antibacterial drugs and their interference with the biogenesis of mitochondria in animal and human cells. Pharm Weekbl Sci. 1983 Jun 24;5(3):81–87. doi: 10.1007/BF01960982. [DOI] [PubMed] [Google Scholar]
  18. Lin C. W., Lin J. C., Prout G. R., Jr Establishment and characterization of four human bladder tumor cell lines and sublines with different degrees of malignancy. Cancer Res. 1985 Oct;45(10):5070–5079. [PubMed] [Google Scholar]
  19. Matlib M. A. Action of bepridil, a new calcium channel blocker on oxidative phosphorylation, oligomycin-sensitive adenosine triphosphatase activity, swelling, Ca++ uptake and Na+-induced Ca++ release processes of rabbit heart mitochondria in vitro. J Pharmacol Exp Ther. 1985 May;233(2):376–381. [PubMed] [Google Scholar]
  20. Moan J., Johannessen J. V., Christensen T., Espevik T., McGhie J. B. Porphyrin-sensitized photoinactivation of human cells in vitro. Am J Pathol. 1982 Nov;109(2):184–192. [PMC free article] [PubMed] [Google Scholar]
  21. Murphy G. F., Shepard R. S., Paul B. S., Menkes A., Anderson R. R., Parrish J. A. Organelle-specific injury to melanin-containing cells in human skin by pulsed laser irradiation. Lab Invest. 1983 Dec;49(6):680–685. [PubMed] [Google Scholar]
  22. Nilsson R., Swanbeck G., Wennersten G. Primary mechanisms of erythrocyte photolysis induced by biological sensitizers and phototoxic drugs. Photochem Photobiol. 1975 Nov;22(5):183–186. doi: 10.1111/j.1751-1097.1975.tb06734.x. [DOI] [PubMed] [Google Scholar]
  23. Park J. K., Dow R. C. The uptake and localization of tetracycline in human blood cells. Br J Exp Pathol. 1970 Apr;51(2):179–182. [PMC free article] [PubMed] [Google Scholar]
  24. Reboud A. M., Dubost S., Reboud J. P. Photoincorporation of tetracycline into rat-liver ribosomes and subunits. Eur J Biochem. 1982 May 17;124(2):389–396. doi: 10.1111/j.1432-1033.1982.tb06605.x. [DOI] [PubMed] [Google Scholar]
  25. Rossi L., Moore G. A., Orrenius S., O'Brien P. J. Quinone toxicity in hepatocytes without oxidative stress. Arch Biochem Biophys. 1986 Nov 15;251(1):25–35. doi: 10.1016/0003-9861(86)90047-0. [DOI] [PubMed] [Google Scholar]
  26. Rømert P., Matthiessen M. E. Swelling of mitochondria in immersion-fixed liver tissue. Effect of various fixatives and of delayed fixation. Acta Anat (Basel) 1981;109(4):332–338. doi: 10.1159/000145399. [DOI] [PubMed] [Google Scholar]
  27. Shea C. R., Wimberly J., Hasan T. Mitochondrial phototoxicity sensitized by doxycycline in cultured human carcinoma cells. J Invest Dermatol. 1986 Sep;87(3):338–342. doi: 10.1111/1523-1747.ep12524430. [DOI] [PubMed] [Google Scholar]
  28. Sorgato M. C., Keller B. U., Stühmer W. Patch-clamping of the inner mitochondrial membrane reveals a voltage-dependent ion channel. Nature. 1987 Dec 3;330(6147):498–500. doi: 10.1038/330498a0. [DOI] [PubMed] [Google Scholar]
  29. Tannock I. F. Oxygen diffusion and the distribution of cellular radiosensitivity in tumours. Br J Radiol. 1972 Jul;45(535):515–524. doi: 10.1259/0007-1285-45-535-515. [DOI] [PubMed] [Google Scholar]
  30. Zuckerman A. J., Baker S. F., Dunkley L. J. The effect of tetracycline on human liver cells in culture. Br J Exp Pathol. 1968 Feb;49(1):20–23. [PMC free article] [PubMed] [Google Scholar]
  31. van den Bogert C., Dontje B. H., Kroon A. M. The antitumour effect of doxycycline on a T-cell leukaemia in the rat. Leuk Res. 1985;9(5):617–623. doi: 10.1016/0145-2126(85)90142-0. [DOI] [PubMed] [Google Scholar]

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