Summary
Treatment with the photosensitizer benzoporphyrin derivative monoacid ring A (BPD-MA, verteporfin) followed by irradiation with visible light induces apoptosis in human acute myelogenous leukaemia HL-60 cells. Photoactivation of BPD-MA induces procaspase 3 (CPP32/Yama/apopain) and procaspase 6 (Mch2) cleavage into their proteolytically active subunits in these cells. The Bcl-2 proto-oncogene product has been shown to protect cells from a number of proapoptotic stimuli. In the present study, the influence of Bcl-2 overexpression on cellular resistance to photoactivation of BPD-MA was studied. Overexpression of Bcl-2 in HL-60 cells prevented apoptosis-related events including caspase 3 and 6 activation, poly(ADP-ribose) polymerase cleavage and the formation of hypodiploid DNA produced by BPD-MA (0–200 ng ml–1) and light. However, Bcl-2 overexpression was less effective at preventing cell death that occurred after photoactivation at high levels (50–100 ng ml–1) compared with lower doses (10–25 ng ml–1) of BPD-MA. These results indicate that caspase 3 and 6 activation and their regulation by Bcl-2 may play important roles in photodynamic therapy (PDT)-induced cell killing.
Similar content being viewed by others
Article PDF
Change history
16 November 2011
This paper was modified 12 months after initial publication to switch to Creative Commons licence terms, as noted at publication
References
Carthy, C. M., Granville, D. J., Watson, K. A., Anderson, D. R., Wilson, J. E., Yang, D. C., Hunt, D. W. C. & McManus, B. M. (1998). Caspase activation and specific cleavage of substrates after coxsackievirus B3-induced cytopathic effect in HeLa cells. J Virol 72: 7669–7675.
Casciola-Rosen, L., Nicholson, D. W., Chong, T., Rowan, K., Thornberry, N. A., Miller, D. & Rosen, A. (1996). Apopain/CPP32 cleaves proteins that are essential for cellular repair: a fundamental principle of apoptotic death. J Exp Med 183: 1957–1964.
Darzynkieicz, Z., Bruno, S., Del Bino, G., Gorczyca, W., Hotz, M., Lassota, P. & Traganos, F. (1992). Features of apoptotic cells measured by flow cytometry. Cytometry 13: 795–808.
Enari, M., Sakahira, H., Yokoyama, H., Okawa, K., Iwamatsu, A. & Nagata, S. (1998). A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 391: 43–50.
Estoppey, S., Rodriguez, I., Sadoul, R. & Martinou, J. C. (1997). Bcl-2 prevents activation of CPP32 cysteine protease and cleavage of poly(ADP-ribose) polymerase and U1-70 kD proteins in staurosporine-mediated apoptosis. Cell Death Differ 4: 34–38.
Fernandes-Alnemri, T., Litwack, G. & Alnemri, E. S. (1995). Mch2, a new member of the apoptotic Ced-3/ICE cysteine protease gene family. Cancer Res 55: 2737–2742.
Gajewski, T. F. & Thompson, C. B. (1996). Apoptosis meets signal transduction: elimination of a BAD influence. Cell 87: 589–593.
Gluck, S., Chadderton, A. & Ho, A. (1996). The selective uptake of benzoporphyrin derivative mono-acid ring A results in differential cell kill of multiple myeloma cells in vitro. Photochem Photobiol 63: 846–853.
Gomer, C. J., Ferrario, A., Hayashi, N., Rucker, N., Szirth, B. & Murphree, A. (1988). Molecular, cellular, and tissue responses following photodynamic therapy. Laser Surg Med 8: 450–463.
Granville, D. J., Levy, J. G. & Hunt, D. W. (1997). Photodynamic therapy induces caspase-3 activation in HL-60 cells. Cell Death Differ 4: 623–629.
Granville, D. J., Levy, J. G. & Hunt, D. W. (1998a). Photodynamic treatment with benzoporphyrin derivative monoacid ring A produces protein tyrosine phosphorylation events and DNA fragmentation in murine P815 cells. Photochem Photobiol 67: 358–362.
Granville, D. J., Jiang, H., An, M. T., Levy, J. G., McManus, B. M. & Hunt, D. W. (1998b). Overexpression of Bcl-X(L) prevents caspase-3-mediated activation of DNA fragmentation factor (DFF) produced by treatment with the photochemotherapeutic agent BPD-MA. FEBS Lett 422: 151–154.
Halenbeck, R., MacDonald, H., Roulston, A., Chen, T. T., Conroy, L. & Williams, L. T. (1998). CPAN, a human nuclease regulated by the caspase-sensitive inhibitor DFF45. Curr Biol 8: 537–540.
He, J., Agarwal, M. L., Larkin, H. E., Friedman, L. R., Xue, L. Y. & Oleinick, N. L. (1996). The induction of partial resistance to photodynamic therapy by the protooncogene BCL-2. Photochem Photobiol 64: 845–852.
Ibrado, A. M., Huang, Y., Fang, G., Liu, L. & Bhalla, K. (1996). Overexpression of Bcl-2 or Bcl-XLinhibits Ara-C-induced CPP32/Yama protease activity and apoptosis of human acute myelogenous leukemia HL-60 cells. Cancer Res 56: 4743–4748.
Ibrado, A. M., Liu, L. & Bhalla, K. (1997). Bcl-XLoverexpression inhibits progression of molecular events leading to paclitaxel-induced apoptosis of human acute myeloid leukemia HL-60 cells. Cancer Res 57: 1109–1115.
Jamieson, C. H. M., McDonald, W. N. & Levy, J. G. (1990). Preferential uptake of benzoporphyrin derivative by leukemic versus normal cells. Leukemia Res 14: 209–219.
Jamieson, C., Richter, A. & Levy, J. (1993). Efficacy of benzoporphyrin derivative, a photosensitizer, in selective destruction of leukemia cells using a murine tumor model. Exp Hematol 21: 629–634.
Kick, G., Messer, G., Plewig, G., Kind, P. & Goetz, A. E. (1995). Strong and prolonged induction of c- junand c- fos proto-oncogenes by photodynamic therapy. Br J Cancer 74: 30–36.
Kluck, R. M., Bossy-Wetzel, E., Green, D. R. & Newmeyer, D. D. (1997). The release of cytochrome c from mitochondria: a primary site for Bcl-2 regulation of apoptosis. Science 275: 1132–1136.
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.
Lazebnik, Y. A., Takahashi, A., Moir, R. D., Goldman, R. D., Poirier, G. G., Kaufmann, S. H. & Earnshaw, W. C. (1995). Studies of the lamin proteinase reveal multiple parallel biochemical pathways during apoptotic execution. Proc Natl Acad Sci USA 92: 9042–9046.
Li, P., Nijhawan, D., Budihardjo, I., Srinivasula, S. M., Ahmad, M., Alnemri, E. S. & Wang, X. (1997). Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 91: 479–489.
Liu, X., Zou, H., Slaughter, C. & Wang, X. (1997). DFF, a heterodimeric protein that functions downstream of caspase-3 to trigger DNA fragmentation during apoptosis. Cell 89: 175–184.
Mosmann, T. (1983). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 57: 55–60.
Nagata, S. (1997). Apoptosis by death factor. Cell 88: 355–365.
Neamati, N., Fernandez, A., Wright, S., Kiefer, J. & McConkey, D. J. (1995). Degradation of Lamin B1 precedes oligonucleosomal DNA fragmentation in apoptotic thymocytes and isolated thymocyte nuclei. J Immunol 154: 3788–3795.
Nicholson, W. D., Ali, A., Thornberry, N. A., Vaillancourt, J. P., Ding, C. K., Gallant, M., Gareau, Y., Griffin, P. R., Labelle, M. & Lazebnik, Y. A. (1995). Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis. Nature 376: 37–43.
Noodt, B. B., Berg, K., Stokke, T., Peng, Q. & Nesland, J. M. (1996). Apoptosis and necrosis induced with light and 5-aminolaevulinic acid-derived protoporphyrin IX. Br J Cancer 74: 22–29.
Orth, K., Chinnaiyan, A. M., Garg, M., Froelich, C. J. & Dixit, V. M. (1996). The CED-3/ICE-like protease Mch2 is activated during apoptosis and cleaves the death substrate lamin A. J Biol Chem 271: 16443–16446.
Pegoraro, L., Palumbo, A., Erikson, J., Falda, M., Giovanazzo, B., Emanuel, B. S., Rovero, G., Nowell, P. C. & Croce, C. M. (1984). A 14;18 and a 8;14 chromosome translocation in a cell line derived from an acute B-cell leukemia. Proc Natl Acad Sci USA 81: 7166–7170.
Perry, D. K., Smyth, M. J., Wang, H. G., Reed, J. C., Poirier, G. G., Obeld, L. M. & Hannun, Y. A. (1997). Bcl-2 acts upstream of the PARP protease and prevents its activation. Cell Death Differ 4: 29–33.
Reed, J. C. (1996). Mechanisms of Bcl-2 family protein function and dysfunction in health and disease. Behring Inst Mitt 97: 72–100.
Reed, J. C. (1997). Double identity for proteins of the Bcl-2 family. Nature 387: 773–776.
Reed, J. C., Miyashita, T., Takayama, S., Wang, H. G., Sato, T., Krajewski, S., Aime-Sempe, C., Bodrug, S., Kitada, S. & Hanada, M. (1996). BCL-2 family proteins: regulators of cell death involved in the pathogenesis of cancer and resistance to therapy. J Cell Biochem 60: 23–32.
Richter, A. M., Kelly, B., Chow, J., Liu, D. J., Towers, G. H., Dolphin, D. & Levy, J. G. (1987). Preliminary studies on a more effective phototoxic agent than hematoporphyrin. J Natl Cancer Inst 79: 1327–1332.
Rowan, S. & Fisher, D. E. (1997). Mechanisms of apoptotic cell death. Leukemia 11: 457–465.
Sakahira, H., Enari, M. & Nagata, S. (1998). Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis. Nature 391: 96–99.
Schlegel, J., Peters, I., Orrenius, S., Miller, D. K., Thornberry, N. A., Yamin, T. & Nicholson, D. W. (1996). CPP32/Apopain is a key interleukin 1b converting enzyme-like protease involved in Fas-mediated apoptosis. J Biol Chem 271: 1841–1844.
Shimizu, S., Eguchi, Y., Kosaka, H., Kamuke, W., Matsuda, H. & Tsujimoto, Y. (1995). Prevention of hypoxia-induced cell death by Bcl-2 and Bcl-XL . Nature 374: 811–813.
Srinivasula, S. M., Fernandes-Alnemri, T., Zangrilli, J., Robertson, N., Armstrong, R. C., Wang, L., Trapani, J. A., Tomaselli, K. J., Litwack, G. & Alnemri, E. S. (1996). The ced-3/interleukin 1beta converting enzyme-like homolog mch6 and the lamin-cleaving enzyme mch2alpha are substrates for the apoptotic mediator CPP32. J Biol Chem 271: 27099–27106.
Tajiri, H., Shinomiya, N., Hayakawa, A., Matsumoto, Y. & Yoshida, S. (1996). Photodynamic therapy-induced rapid cell death by apoptosis in human pancreatic carcinoma transplanted into nude mice. J Clin Biochem Nutr 21: 29–37.
Takahashi, A., Alnemri, E. S., Lazebnik, Y. A., Fernandes-Alnemri, T., Litwack, G., Moir, R. D., Goldman, R. D., Poirier, G. G., Kaufmann, S. H. & Earnshaw, W. C. (1996). Cleavage of lamin A by Mch2a but not CPP32: multiple interleukin 1b-converting enzyme-related proteases with distinct substrate recognition properties are active in apoptosis. Proc Natl Acad Sci USA 93: 8395–8400.
Telford, W. G., King, L. E. & Fraker, P. J. (1994). Rapid quantitation of apoptosis in pure and heterogenous cell populations using flow cytometry. J Immunol Methods 172: 1
Thornberry, N. A., Rano, T. A., Peterson, E. P., Rasper, D. M., Timkey, T., Garcia-Calvo, M., Houtzager, V. M., Nordstrom, P. A., Roy, S., Vaillancourt, J. P., Chapman, K. T. & Nicholson, D. W. (1997). A combinatorial approach defines specificities of members of the caspase family and granzyme B. Functional relationships established for key mediators of apoptosis. J Biol Chem 272: 17907–17911.
Tsujimoto, Y., Cossman, J., Jaffe, E. & Croce, C. M. (1985). Involvement of the bcl-2 gene in human follicular lymphoma. Science 228: 1440–1443.
Wang, X., Zelenski, N. G., Yang, J., Sakai, J., Brown, M. S. & Goldstein, J. L. (1996). Cleavage of sterol regulatory element binding proteins (SREBPs) by CPP32 during apoptosis. EMBO J 15: 1012–1020.
Yang, E., Zha, J., Jockel, J., Boise, L., Thompson, C. B. & Korsemeyer, S. J. (1995). Bad, a heterodimeric partner for Bcl-XLand Bcl-2, displaces Bax and promotes cell death. Cell 80: 285–291.
Yang, J., Liu, X., Bhalla, K., Kim, C. N., Ibrado, A. M., Cai, J., Peng, T., Jones, D. P. & Wang, X. (1997). Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science 275: 1129–1132.
Zaidi, S. I., Oleinick, N. L., Zaim, M. T. & Mukhtar, H. (1993). Apoptosis during photodynamic therapy-induced ablation of RIF-1 tumors in C3H mice: electron microscopic, histopathologic and biochemical evidence. Photochem Photobiol 58: 771–776.
Author information
Authors and Affiliations
Rights and permissions
From twelve months after its original publication, this work is licensed under the Creative Commons Attribution-NonCommercial-Share Alike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
About this article
Cite this article
Granville, D., Jiang, H., An, M. et al. Bcl-2 overexpression blocks caspase activation and downstream apoptotic events instigated by photodynamic therapy. Br J Cancer 79, 95–100 (1999). https://doi.org/10.1038/sj.bjc.6690017
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.bjc.6690017
Keywords
This article is cited by
-
p38 MAPK plays an essential role in apoptosis induced by photoactivation of a novel ethylene glycol porphyrin derivative
Oncogene (2008)
-
Heme oxygenase-1 protects tumor cells against photodynamic therapy-mediated cytotoxicity
Oncogene (2006)
-
Apoptosis of gastric cancer cell line MKN45 by photodynamic treatment with Photofrin
Lasers in Medical Science (2004)
-
Bax is essential for mitochondrion-mediated apoptosis but not for cell death caused by photodynamic therapy
British Journal of Cancer (2003)
-
Photodamage to multiple Bcl-xL isoforms by photodynamic therapy with the phthalocyanine photosensitizer Pc 4
Oncogene (2003)