Photodynamic Therapy and Cell Death Pathways

  • David Kessel
  • Nancy L. Oleinick
Part of the Methods in Molecular Biology book series (MIMB, volume 635)


Photodynamic therapy (PDT) is the term used to describe the irradiation of photosensitized cells or tissue with phototoxic consequences. This process can result in the rapid initiation of not only apoptosis, an irreversible death pathway, but also autophagy. The procedures described here are designed to characterize the correlation between the PDT dose vs. survival of cells in vitro, the apoptotic effects of photodamage, and the extent of an autophagic response. These are assessed by clonogenic assays, observation of condensed chromatin characteristic of apoptosis, activation of “executioner” caspases, and the autophagic flux as indicated by comparing accumulation of the LC3-II protein under conditions where processing of autophagosomes is retarded vs. is not retarded.

Key words

Apoptosis autophagy Bcl-2 Bid Bax photosensitization photodynamic therapy 



The authors’ research is supported by NIH grant CA 23378 (to DK) and NIH grant CA 106491 (to NLO) from the National Cancer Institute, DHHS and by the State of Ohio Biomedical Research and Technology Transfer Trust TECH 05-063 (to NLO).


  1. 1.
    Dougherty, T. J., Gomer, C. J., Henderson, B. W., Jori, G., Kesse, L. D., Korbelik, M., Moan, J., and Peng, Q. (1998) Photodynamic therapy. J Natl Cancer Inst, 90, 889–905.PubMedCrossRefGoogle Scholar
  2. 2.
    Kim, H. R., Luo, Y., Li, G., and Kessel, D. (1999) Enhanced apoptotic response to photodynamic therapy after Bcl-2 transfection. Cancer Res, 59, 3429–3432.PubMedGoogle Scholar
  3. 3.
    Kessel, D. and Castelli, M. (2001) Evidence that Bcl-2 is the target of three photosensitizers that induce a rapid apoptotic response. Photochem Photobiol, 74, 318–322.PubMedCrossRefGoogle Scholar
  4. 4.
    Xue, L. Y., Chiu, S. M., and Oleinick, N. L. (2001) Photochemical destruction of the Bcl-2 oncoprotein during photodynamic therapy with the phthalocyanine photosensitizer Pc 4. Oncogene, 20, 3420–3427.PubMedCrossRefGoogle Scholar
  5. 5.
    Xue, L. Y., Chiu, S. M., Fiebig, A., Andrews, D. W., and Oleinick, N. L. (2003) Photodamage to multiple Bcl-xL isoforms by photodynamic therapy with the phthalocyanine photosensitizer Pc 4. Oncogene, 22, 9197–9204.PubMedCrossRefGoogle Scholar
  6. 6.
    Kessel, D. and Reiners, J. J., Jr. (2007) Apoptosis and autophagy after mitochondrial or endoplasmic reticulum photodamage. Photochem Photobiol, 83, 1024–1028.PubMedCrossRefGoogle Scholar
  7. 7.
    Reiners, J. J., Jr., Caruso., J. A., Mathieu, P., Chelladurai, B., Yin, X. M., and Kessel, D. (2002) Release of cytochrome c and activation of pro-caspase-9 following lysosomal photodamage involves Bid cleavage. Cell Death Differ, 9, 934–944.PubMedCrossRefGoogle Scholar
  8. 8.
    Rodriguez, M. E., Azizuddin, K., Chiu, S. M., Xue, L. Y., Zhang, P., Lam, M., Kenney, M. E., Nieminen, A. L., and Oleinick, N. L. (2009) Structural factors and mechanisms underlying the improved photodynamic cell killing with silicon phthalocyanine photosensitizers directed to lysosomes vs. mitochondria. Photochem Photobiol, 85, 1189–1200.PubMedCrossRefGoogle Scholar
  9. 9.
    Buytaert, E., Callewaert, G., Hendrickx, N., Scorrano, L., Hartmann, D., Missiaen, L., Vandenheede, J. R., Heirman, I., Grooten, J., and Agostinis, P. (2006) Role of endoplasmic reticulum depletion and multidomain proapoptotic BAX and BAK proteins in shaping cell death after hypericin-mediated photodynamic therapy. FASEB J, 20, 756–758.PubMedGoogle Scholar
  10. 10.
    Kessel, D. and Arroyo, A. S. (2007) Apoptotic and autophagic responses to Bcl-2 inhibition and photodamage. Photochem Photobiol Sci, 6, 1290–1295.PubMedCrossRefGoogle Scholar
  11. 11.
    Kessel, D., Vicente, M. G., and Reiners, J. J., Jr. (2006) Initiation of apoptosis and autophagy by photodynamic therapy. Lasers Surg Med, 38, 482–488.PubMedCrossRefGoogle Scholar
  12. 12.
    Xue, L. Y., Chiu, S. M., Azizuddin, K., Joseph, S., and Oleinick, N. L. (2007) The death of human cancer cells following photodynamic therapy: apoptosis competence is necessary for Bcl-2 protection but not for induction of autophagy. Photochem Photobiol, 83, 1016–1023.PubMedCrossRefGoogle Scholar
  13. 13.
    Klionsky, D. et al. (2008) Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes. Autophagy, 4, 151–175.PubMedGoogle Scholar
  14. 14.
    Mizushima, N. and Yoshimori, T. (2007) How to interpret LC3 immunoblotting. Autophagy, 3, 542–545.PubMedGoogle Scholar
  15. 15.
    Pryde, J. G., Walker, A., Rossi, A. G., Hannah, S., and Haslett, C. (2000) Temperature-dependent arrest of neutrophil apoptosis. Failure of Bax insertion into mitochondria at 15 degrees C prevents the release of cytochrome c. J Biol Chem, 275, 33574–33584.PubMedCrossRefGoogle Scholar
  16. 16.
    Iwai-Kanai, E., Yuan, H., Huang, C., Sayen, M. R., Perry-Garza, C. N., Kim, L., and Gottlieb, R. A. (2008) A method to measure cardiac autophagic flux in vivo. Autophagy, 4, 322–329.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • David Kessel
    • 1
  • Nancy L. Oleinick
    • 2
  1. 1.Wayne State University School of MedicineDetroitUSA
  2. 2.Case Western Reserve UniversityClevelandUSA

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