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  • Original Paper
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Photochemical destruction of the Bcl-2 oncoprotein during photodynamic therapy with the phthalocyanine photosensitizer Pc 4

Abstract

Photodynamic therapy (PDT), utilizing a photosensitizer and visible light, causes localized oxidative damage. With the mitochondrial photosensitizer Pc 4, PDT induces apoptosis, yet its molecular targets are not known. Here, the anti-apoptotic protein Bcl-2 is shown to be highly sensitive to PDT, as judged on Western blots by the disappearance of anti-Bcl-2-reactive material from the position of the native 26 kDa protein. The loss of Bcl-2 was PDT dose dependent and was observed for both endogenous and overexpressed Bcl-2 in several cell lines, immediately after PDT, and with chilled cells. It was accompanied by a trace of a 23-kDa cleavage product as well as high-molecular weight products that may result from photochemical crosslinking. PDT-induced Bcl-2 loss occurred in MCF-7 cells that do not express caspase-3 or in the presence of protease inhibitors, but was prevented, along with the induction of apoptosis, by the singlet oxygen scavenger L-histidine. Loss of FLAG-Bcl-2 was observed with both anti-FLAG and anti-Bcl-2 antibodies, indicating loss of native protein rather than simple BCL-2-epitope destruction. Photochemical damage was not observed in Bcl-xL, Bax, Bad, the voltage-dependent anion channel, or the adenine nucleotide translocator. Therefore, Bcl-2 is one target of PDT with Pc 4, and PDT damage to Bcl-2 contributes to its efficient induction of apoptosis.

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References

  • Agarwal R, Athar M, Urban SA, Bickers DR, Mukhtar H . 1991 Cancer Lett. 56: 125–129

  • Antonsson B, Conti F, Ciavatta A, Montessuit S, Lewis S, Martinou I, Bernasconi L, Bernard A, Mermod JJ, Mazzei G, Maundrell K, Gambale F, Sadoul R, Martinou JC . 1997 Science 277: 370–372

  • Bissonnette RP, Echeverri F, Mahboubi A, Green DR . 1992 Nature 359: 552–554

  • Carthy CM, Granville DJ, Jiang H, Levy JG, Rudin CM, Thompson CB, McManus BM, Hunt DW . 1999 Lab. Invest. 79: 953–965

  • Cheng EH, Kirsch DG, Clem RJ, Ravi R, Kastan MB, Bedi A, Ueno K, Hardwick JM . 1997 Science 278: 1966–1968

  • Colussi VC, Feyes DK, Mulvihill JW, Li YS, Kenney ME, Elmets CA, Oleinick NL, Mukhtar H . 1999 Photochem. Photobiol. 69: 236–241

  • Eguchi Y, Ewert DL, Tsujimoto Y . 1992 Nucleic Acids Res. 20: 4187–4192

  • Fadeel B, Hassan Z, Hellstrom-Lindberg E, Henter JI, Orrenius S, Zhivotovsky B . 1999 Leukemia 13: 719–728

  • Fujita N, Tsuruo T . 1998 Biochem. Biophys. Res. Commun. 246: 484–488

  • Grandgirard D, Studer E, Monney L, Belser T, Fellay I, Borner C, Michel MR . 1998 EMBO J. 17: 1268–1278

  • Granville DJ, Jiang H, An MT, Levy JG, McManus BM, Hunt DW . 1999 Br. J. Cancer 79: 95–100

  • Gross A, McDonnell JM, Korsmeyer SJ . 1999 Genes Dev. 13: 1899–1911

  • Haldar S, Chintapalli J, Croce CM . 1996 Cancer Res. 56: 1253–1255

  • Halliwell B, Gutteridge JMC . 1989 Free Radicals in Biology and Medicine, 2nd edition Oxford University Press: Oxford pp. 66–70

  • He H, Qi XM, Grossmann J, Distelhorst CW . 1998 J. Biol. Chem. 273: 25015–25019

  • He J, Agarwal ML, Larkin HE, Friedman LR, Xue L-Y, Oleinick NL . 1996 Photochem. Photobiol. 64: 845–852

  • He J, Larkin HE, Li Y-S, Rihter BD, Zaidi SIA, Rodgers MAJ, Mukhtar H, Kenney ME, Oleinick NL . 1997 Photochem. Photobiol. 65: 581–586

  • Henderson BW, Dougherty TJ . 1992 Photochem. Photobiol. 55: 145–157

  • Hockenbery DM, Oltvai ZN, Yin XM, Milliman CL, Korsmeyer SJ . 1993 Cell 75: 241–251

  • Jacobsen MD, Weil M, Raff MC . 1996 J. Cell Biol. 133: 1041–1051

  • Janicke RU, Sprengart ML, Wati MR, Porter AG . 1998 J. Biol. Chem. 273: 9357–9360

  • Kessel D, Luo Y . 1999 Cell Death Differ. 6: 28–35

  • Kim HR, Luo Y, Li G, Kessel D . 1999 Cancer Res. 59: 3429–3432

  • Kirsch DG, Doseff A, Chau BN, Lim DS, de Souza-Pinto NC, Hansford R, Kastan MB, Lazebnik YA, Hardwick JM . 1999 J. Biol. Chem. 274: 21155–21161

  • Kluck RM, Bossy-Wetzel E, Greene DR, Newmeyer DW . 1997 Science 275: 1132–1136

  • Korsmeyer SJ . 1999 Cancer Res. 59: 1693s–1700s

  • Lam M, Dubyak G, Chen L, Nunez G, Miesfeld RL, Distelhorst CW . 1994 Proc. Natl. Acad. Sci. USA 91: 6569–6573

  • Lavie G, Kaplinsky C, Toren A, Aizman I, Meruelo D, Mazur Y, Mandel M . 1999 Br. J. Cancer 79: 423–432

  • Moan J, Berg K . 1992 Photochem. Photobiol. 55: 931–948

  • Oleinick NL, Antunez AR, Clay ME, Rihter BD, Kenney ME . 1993 Photochem. Photobiol. 57: 242–247

  • Oleinick NL, Evans HH . 1998 Radiat. Res. 150: S146–S156

  • Qiu Y, Robinson D, Pretlow TG, Kung HJ . 1998 Proc. Natl. Acad. Sci. USA 95: 3644–3649

  • Reed JC, Jurgensmeier JM, Matsuyama S . 1998 Biochim. Biophys. Acta 1366: 127–137

  • Susin SA, Lorenzo HK, Zamzami N, Marzo I, Brenner C, Larochette N, Prevost MC, Alzari PM, Kroemer G . 1999 J. Exp. Med. 189: 381–394

  • Trivedi NS, Wang HW, Nieminen AL, Oleinick NL, Izatt JA . 2000 Photochem. Photobiol. 71: 634–639

  • Tsujimoto Y, Croce CM . 1986 Proc. Natl. Acad. Sci. USA 83: 5214–5218

  • Tudor G, Aguilera A, Halverson DO, Laing ND, Sausville EA . 2000 Cell Death Differ. 7: 574–586

  • Varnes ME, Chiu SM, Xue LY, Oleinick NL . 1999 Biochem. Biophys. Res. Commun. 255: 673–679

  • Weishaupt KR, Gomer CJ, Dougherty TJ . 1976 Cancer Res. 36: 2326–2329

  • Wuerzberger SM, Pink JJ, Planchon SM, Byers KL, Bornmann WG, Boothman DA . 1998 Cancer Res. 58: 1876–1885

  • Xue LY, Chiu SM, Oleinick NL . 2001 Exp. Cell Res. 263: 145–155

  • Xue LY, Qiu Y, He J, Kung HJ, Oleinick NL . 1999 Oncogene 18: 3391–3398

  • Yang J, Liu X, Bhalla K, Kim CN, Ibrado AM, Cai J, Peng TI, Jones DP, Wang X . 1997 Science 275: 1129–1132

  • Zhang WG, Ma LP, Wang SW, Zhang ZY, Cao GD . 1999 Photochem. Photobiol. 69: 582–586

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Acknowledgements

The authors thank Drs Christopher J Froelich, David A Boothman, John J Pink and Sarah M Planchon for providing some of the cell lines used in this study. This research was supported by U.S. Public Health Service Grants P01 CA48735, R01 CA83917, and P30 CA43703 from the National Cancer Institute, DHHS.

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Xue, Ly., Chiu, Sm. & Oleinick, N. Photochemical destruction of the Bcl-2 oncoprotein during photodynamic therapy with the phthalocyanine photosensitizer Pc 4. Oncogene 20, 3420–3427 (2001). https://doi.org/10.1038/sj.onc.1204441

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