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Involvement of both caspase-dependent and -independent pathways in apoptotic induction by hexaminolevulinate-mediated photodynamic therapy in human lymphoma cells

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Abstract

Photodynamic therapy (PDT) is a cancer treatment based on the interaction of a photosensitizer, light and oxygen. PDT with the endogenous photosensitizer, protoporphyrin IX (PpIX) induced by 5-aminolevulinic acid (ALA) or its derivatives is a modification of this treatment modality with successful application in dermatology. However, the mechanism of cell destruction by ALA-PDT has not been elucidated. In this study a human T-cell lymphoma Jurkat cell line was treated with PDT using hexaminolevulinate (HAL, hexylester of ALA). Four hours following treatment nearly 80% of the cells exhibited typical apoptotic features. Mitochondrial pro-apoptotic proteins were evaluated by Western blots in subcellular fractionated samples. PDT caused cytosolic translocation of cytochrome c and nuclear redistribution of apoptosis-inducing factor (AIF), but the release of mitochondrial Smac/DIABLO, Omi/HtrA2 and EndoG was not observed. The release of cytochrome c was followed by the cleavage of caspase-9 and caspase-3 as well as its downstream substrates, together with oligonucleosomal DNA fragmentation. The pan-caspases inhibitor, z-VAD.fmk, prevented oligonucleosomal DNA fragmentation, but failed to inhibit PDT-mediated apoptosis. The apoptotic induction by AIF-mediated caspase-independent pathway was also found after HAL-PDT with large-scale DNA fragmentation in the presence of z-VAD.fmk. These results demonstrate that cytochrome c-mediated caspase-dependent pathway and AIF-induced caspase-independent pathway are simultaneously involved in the apoptotic induction by PDT. When the cytochrome c-induced caspase-dependent pathway is blocked, the cells go into apoptosis via AIF-mediated pathway, clearly demonstrating that the cytochrome c-mediated caspase-dependent pathway is not required for such apoptotic induction. This finding may have an impact on improved PDT effectiveness.

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Authors and Affiliations

  1. Pathology Clinic, Rikshospitalet-Radiumhospitalet HF Medical Center, Faculty Division Radiumhospitalet, University of Oslo, Norway

    Ingegerd Eggen Furre, Michael T. N. Møller, Susan Shahzidi, Jahn M. Nesland & Qian Peng

  2. State Key Lab for Advanced Photonic Materials and Device, Fudan University, Shanghai, 200433, China

    Qian Peng

  3. Department of Pathology, The Norwegian Radium Hospital, University of Oslo, Montebello, 0310, Oslo, Norway

    Qian Peng

Authors
  1. Ingegerd Eggen Furre
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  2. Michael T. N. Møller
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  3. Susan Shahzidi
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  4. Jahn M. Nesland
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  5. Qian Peng
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Correspondence to Qian Peng.

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Furre, I.E., Møller, M.T.N., Shahzidi, S. et al. Involvement of both caspase-dependent and -independent pathways in apoptotic induction by hexaminolevulinate-mediated photodynamic therapy in human lymphoma cells. Apoptosis 11, 2031–2042 (2006). https://doi.org/10.1007/s10495-006-0190-x

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  • DOI: https://doi.org/10.1007/s10495-006-0190-x

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