Abstract
Photodynamic therapy (PDT) can result in both types of cell death, apoptosis or necrosis. Several steps in the induction and execution of apoptosis depend on ATP and the intracellular ATP level has been shown to be one determinant in whether apoptosis or necrosis occurs. Therefore, photochemical damage of cellular targets involved in energy supply might play a crucial role in the mode of cell death being executed. The present study is aimed at the characterization of changes in cellular energy supply and the associated cell death modes in response to PDT. Using the human epidermoid carcinoma cell line A431 and aluminium(iii) phthalocyanine tetrasulfonate chloride (2.5 μM) as a photosensitizer, we studied the changes in mitochondrial function and intracellular ATP level after irradiation with different light doses. Employing assays for caspase-3 activation and nuclear fragmentation, 50of the cells were found to undergo apoptosis after irradiation between 2.5 to 3.5 J cm−2 while the remainder died by necrosis. At higher light doses (>6 J cm−2), neither caspase-3 activation nor nuclear fragmentation was observed and this suggests that these cells died exclusively by necrosis. Necrotic cell death was also associated with a rapid decline in mitochondrial activity and intracellular ATP. By contrast, with apoptosis the loss of mitochondrial function was delayed and the ATP level was maintained at near control levels for up to eight hours which was far beyond the onset of morphological changes. These data suggest that, depending on the light dose applied, both, necrosis as well as apoptosis can be induced with AlPcS4 mediated PDT and that photodamage in energy supplying cellular targets may influence the mode of cell death. Further, it is speculated that cells undergoing apoptosis in response to PDT might maintain a high ATP level long enough to complete the apoptotic program.
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References
T. J. Dougherty, Photodynamic therapy, Photochem. Photobiol., 1993, 58(6), 895–900.
B. W. Henderson and T. J. Dougherty, How does photodynamic therapy work?, Photochem. Photobiol, 1992, 55(1), 145–157.
A. Strasser, L. O’Connor and V. M. Dixit, Apoptosis signaling, Annu. Rev. Biochem., 2000, 69, 217–245.
N. L. Oleinick, R. L. Morris and I. Belichenko, The role of apoptosis in response to photodynamic therapy: what, where, why, and how, Photochem. Photobiol. Sci., 2002, 1(1), 1–21.
I. Reiter, B. Krammer and G. Schwamberger, Cutting edge: differential effect of apoptotic versus necrotic tumor cells on macrophage antitumor activities, J. Immunol., 1999, 163(4), 1730–2.
V. A. Fadok, D. L. Bratton, L. Guthrie and P. M. Henson, Differential effects of apoptotic versus lysed cells on macrophage production of cytokines: role of proteases, J. Immunol., 2001, 166(11), 6847–6854.
B. Sauter, M. L. Albert, L. Francisco, M. Larsson, S. Somersan and N. Bhardwaj, Consequences of cell death: exposure to necrotic tumor cells, but not primary tissue cells or apoptotic cells, induces the maturation of immunostimulatory dendritic cells, J. Exp. Med., 2000, 191(3), 423–434.
A. C. Moor, Signaling pathways in cell death and survival after photodynamic therapy, J. Photochem. Photobiol. B, 2000, 57(1), 1–13.
G. M. Malham, R. J Thomsen, G. J. Finlay and B. C. Baguley, Subcellular distribution and photocytotoxicity of aluminium phthalocyanines and haematoporphyrin derivative in cultured human meningioma cells, Br. J. Neurosurg., 1996, 10(1), 51–57.
L. Wyld, M. W. Reed and N. J. Brown, Differential cell death response to photodynamic therapy is dependent on dose and cell type, Br. J. Cancer, 2001, 84(10), 1384–1386.
A. Villanueva, V. Dominguez, S. Polo, V. D. Vendrell, C. Sanz, T. M. Canete, A. Juarranz and J. C. Stockert, Photokilling mechanisms induced by zinc(II)-phthalocyanine on cultured tumor cells, Oncol. Res., 1999, 11(10), 447–453.
G. Lavie, C. Kaplinsky, A. Toren, I. Aizman, D. Meruelo, Y. Mazur and M. Mandel, A photodynamic pathway to apoptosis and necrosis induced by dimethyl tetrahydroxyhelianthrone and hypericin in leukaemic cells: possible relevance to photodynamic therapy, Br. J. Cancer, 1999, 79(3–4), 423–432.
Y. Luo and D. Kessel, Initiation of apoptosis versus necrosis by photodynamic therapy with chloroaluminum phthalocyanine, Photochem. Photobiol., 1997, 66(4), 479–483.
B. B. Noodt, G. H. Rodal, M. Wainwright, Q. Peng, R. Horobin, J. M. Nesland and K. Berg, Apoptosis induction by different pathways with methylene blue derivative and light from mitochondrial sites in V79 cells, Int. J. Cancer, 1998, 75(6), 941–948.
C. Richter, M. Schweizer, A. Cossarizza and C. Franceschi, Control of apoptosis by the cellular ATP level, FEBS Lett., 1996, 378(2), 107–110.
P. Nicotera, M. Leist and E. FerrandoMay, Intracellular ATP, a switch in the decision between apoptosis and necrosis, Toxicol. Lett., 1998, 103, 139–142.
M. Leist, B. Single, A. F. Castoldi, S. Kuhnle and P. Nicotera, Intracellular adenosine triphosphate (ATP) concentration: A switch in the decision between apoptosis and necrosis, J. Exp. Med., 1997, 185(8), 1481–1486.
Y. Eguchi, S. Shimizu and Y. Tsujimoto, Intracellular ATP levels determine cell death fate by apoptosis or necrosis, Cancer Res., 1997, 57(10), 1835–1840.
Y. Hu, M. A. Benedict, L. Ding and G. Nunez, Role of cytochrome c and dATP/ATP hydrolysis in Apaf-1-mediated caspase-9 activation and apoptosis, EMBO J., 1999, 18(13), 3586–3595.
Y. Eguchi, A. Srinivasan, K. J. Tomaselli, S. Shimizu and Y. Tsujimoto, ATP-dependent steps in apoptotic signal transduction, Cancer Res., 1999, 59(9), 2174–2181.
G. E. Kass, J. E. Eriksson, M. Weis, S. Orrenius and S. C. Chow, Chromatin condensation during apoptosis requires ATP, Biochem. J., 1996, 318(Pt 3), 749–752.
A. Saleh, S. M. Srinivasula, S. Acharya, R. Fishel and E. S. Alnemri, Cytochrome c and dATP-mediated oligomerization of Apaf-1 is a prerequisite for procaspase-9 activation, J. Biol. Chem., 1999, 274(25), 17941–17945.
D. Kessel and Y. Luo, Mitochondrial photodamage and PDT-induced apoptosis, J. Photochem. Photobiol. B, 1998, 42(2), 89–95.
V. Kirveliene, L. Prasmickaite, J. Kadziauskas, R. Bonnett, B. D. Djelal and B. Juodka, Post-exposure processes in Temoporfin-photosensitized cells in vitro: reliance on energy metabolism, J. Photochem. Photobiol. B, 1997, 41(1–2), 173–180.
T. Mosmann, Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays, J. Immunol. Methods, 1983, 65(1–2), 55–63.
D. W. Nicholson, Caspase structure, proteolytic substrates, and function during apoptotic cell death, Cell Death Differ., 1999, 6(11), 1028–1042.
W. C. Earnshaw, L. M. Martins and S. H. Kaufmann, Mammalian caspases: structure, activation, substrates, and functions during apoptosis, Annu. Rev. Biochem., 1999, 68, 383–424.
D. J. Ball, S. Mayhew, D. I. Vernon, M. Griffin and S. B. Brown, Decreased efficiency of trypsinization of cells following photodynamic therapy: evaluation of a role for tissue transglutaminase, Photochem. Photobiol., 2001, 73(1), 47–53.
H. A. Neufeld, R. D. Towner and J. Pace, A rapid method for determining ATP by the firefly luciferin-luciferase system, Experientia, 1975, 31(3), 391–392.
G. Marcaida, M. D. Minana, S. Grisolia and V. Felipo, Determination of intracellular ATP in primary cultures of neurons, Brain Res. Brain Res. Protoc., 1997, 1(1), 75–78.
A. Ruck, K. Heckelsmiller, R. Kaufmann, N. Grossman, E. Haseroth and N. Akgun, Light-induced apoptosis involves a defined sequence of cytoplasmic and nuclear calcium release in AlPcS4-photosensitized rat bladder RR 1022 epithelial cells, Photochem. Photobiol., 2000, 72(2), 210–216.
G. Jori and C. Fabris, Relative contributions of apoptosis and random necrosis in tumour response to photodynamic therapy: effect of the chemical structure of Zn(II)-phthalocyanines, J. Photochem. Photobiol. B, 1998, 43(3), 181–185.
J. Cai, J. Yang and D. Jones, Mitochondrial control of apoptosis: the role of cytochrome c, Biochim. Biophys. Acta, 1998, 1366(1–2), 139–149.
S. A. Susin, H. K. Lorenzo, N. Zamzami, I. Marzo, B. E. Snow, G. M. Brothers, J. Mangion, E. Jacotot, P. Costantini, M. Loeffler, N. Larochette, D. R. Goodlett, R. Aebersold, D. P. Siderovski, J. M. Penninger and G. Kroemer, Molecular characterization of mitochondrial apoptosis-inducing factor, Nature, 1999, 397(6718), 441–446.
D. Kulms and T. Schwarz, Molecular mechanisms of UV-induced apoptosis, Photodermatol. Photoimmunol. Photomed., 2000, 16(5), 195–201.
C. A. Belmokhtar, J. Hillion, E. Segal-Bendirdjian, Staurosporine induces apoptosis through both caspase-dependent and caspase-independent mechanisms, Oncogene, 2001, 20(26), 3354–3362.
R. Bertrand, E. Solary, P. O’Connor, K. W. Kohn and Y. Pommier, Induction of a common pathway of apoptosis by staurosporine, Exp. Cell Res., 1994, 211(2), 314–321.
Z. Han, P. Pantazis, T. S. Lange, J. H. Wyche and E. A. Hendrickson, The staurosporine analog, Ro-31-8220, induces apoptosis independently of its ability to inhibit protein kinase C, Cell Death Differ., 2000, 7(6), 521–530.
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Plaetzer, K., Kiesslich, T., Krammer, B. et al. Characterization of the cell death modes and the associated changes in cellular energy supply in response to AlPcS4-PDT. Photochem Photobiol Sci 1, 172–177 (2002). https://doi.org/10.1039/b108816e
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DOI: https://doi.org/10.1039/b108816e