Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

Photodamage to multiple Bcl-xL isoforms by photodynamic therapy with the phthalocyanine photosensitizer Pc 4

Oncogene volume 22, pages 9197–9204 (2003)Cite this article

Abstract

The antiapoptotic oncoprotein Bcl-2 is now a recognized phototarget of photodynamic therapy (PDT) with the phthalocyanine Pc 4 and with other mitochondrion-targeting photosensitizers. Photodamage, observed on Western blots as the loss of the native 26-kDa Bcl-2 protein, is PDT dose dependent and occurs in multiple cell lines, in the cold, and immediately upon photoirradiation. In our initial study, no photochemical damage was observed to Bcl-xL, in spite of its similarity in size, sequence, location and function to Bcl-2. The original study used a commercial anti-Bcl-xS/L antibody. We have revisited this issue by examining Western blots developed using one of three epitope-specific anti-Bcl-xL antibodies from commercial sources, a polyclonal antibody generated to the entire protein, as well as the antibody used previously. All five Bcl-xL antibodies recognized bacterially expressed Bcl-xL, but not Bcl-2, whereas an anti-Bcl-2 antibody recognized Bcl-2 and not Bcl-xL. All five Bcl-xL antibodies recognized at least one protein migrating at ∼30 kDa; two of the antibodies recognized an additional band, migrating at ∼33 or ∼24 kDa. We now observe Pc 4-PDT-induced photodamage to all Bcl-xL-related proteins, except the 33-kDa species, in several human cancer cell lines. The results indicate that, in addition to the expected quantitative differences that may reflect exposure of individual epitopes, the antibodies also detect proteins of different apparent molecular weights that may be distinct isoforms or post-translationally modified forms of Bcl-xL. No evidence for PDT-induced phosphorylation or degradation was observed. Bcl-xL localized to mitochondria was considerably more sensitive to photodamage than was Bcl-xL in the cytosol, indicating that as previously found for Bcl-2, Bcl-xL must be membrane localized to be photosensitive.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  • Boise LH, Gonzalez-Garcia M, Postema CE, Ding L, Lindsten T, Turka LA, Mao X, Nunez G and Thompson CB . (1993). Cell, 74, 597–608.

  • Chao DT and Korsmeyer SJ . (1998). Annu. Rev. Immunol., 16, 395–419.

  • Chao DT, Linette GP, Boise LH, White LS, Thompson CB and Korsmeyer SJ . (1995). J. Exp. Med., 182, 821–828.

  • Dougherty TJ, Gomer CJ, Henderson BW, Jori G, Kessel D, Korbelik M, Moan J and Peng Q . (1998). J. Natl. Cancer Inst., 90, 889–905.

  • Fang W, Rivard J J, Mueller D L and Behrens W . (1994). J. Immunol., 153, 4388–4398.

  • Gonzalez-Garcia M, Perez-Ballestero R, Ding L, Duan L, Boise LH, Thompson CB and Nunez G . (1994). Development, 120, 3033–3042.

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

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

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

  • Hockenbery D, Nunez G, Milliman C, Schreiber RD and Korsmeyer SJ . (1990). Nature, 348, 334–336.

  • Hsu YT and Youle RJ . (1997). J. Biol. Chem., 272, 13829–13834.

  • Kessel D and Castelli M . (2001). Photochem. Photobiol., 74, 318–322.

  • Kessel D and Luo Y . (1999). Cell Death Differ., 6, 28–35.

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

  • Krajewski S, Tanaka S, Takayama S, Schibler MJ, Fenton W and Reed JC . (1993). Cancer Res., 53, 4701–4714.

  • Kroemer G . (1997). Nat. Med., 3, 614–620.

  • Lam M, Oleinick NL and Nieminen AL . (2001). J. Biol. Chem., 276, 47379–47386.

  • Li X, Marani M, Mannucci R, Kinsey B, Andriani F, Nicoletti I, Denner L and Marcelli M . (2001). Cancer Res., 61, 1699–1706.

  • Liu QY and Stein CA . (1997). Clin. Cancer Res., 3, 2039–2046.

  • Moan J and Berg K . (1991). Photochem. Photobiol., 53, 549–553.

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

  • Oleinick NL and Evans HH . (1998). Radiat. Res., 150, S146–S156.

  • Oleinick NL, Morris RL and Belichenko I . (2002). Photochem. Photobiol. Sci., 1, 1–21.

  • Reed JC . (1995). Hematol. Oncol. Clin. N. Am., 9, 451–473.

  • Srivastava M, Ahmad N, Gupta S and Mukhtar H . (2001). J. Biol. Chem., 276, 15481–15488.

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

  • Usuda J, Azizuddin K, Chiu SM and Oleinick NL . (2003a). Photochem. Photobiol., 78, 1–8.

  • Usuda J, Chiu SM, Murphy ES, Lam M, Nieminen AL and Oleinick NL . (2003b). J. Biol. Chem., 278, 2021–2029.

  • Wang NS, Unkila MT, Reineks EZ and Distelhorst CW . (2001). J. Biol. Chem., 276, 44117–44128.

  • White E . (1996). Genes Dev., 10, 1–15.

  • Xue LY, Chiu SM and Oleinick NL . (2001). Oncogene, 20, 3420–3427.

  • Ye Q, Press B, Kissler S, Yang X, Lu L, Bassing CH, Sleckman BP, Jansson M, Panoutsakopoulou V, Trimble LA, Alt FW and Cantor H . (2002). J. Exp. Med., 196, 87–95.

Download references

Acknowledgements

We thank Dr Clark W Distelhorst, Departments of Medicine and Pharmacology, CWRU, for the bacterial cultures expressing Bcl-2 and Bcl-xL, and Drs Sarah M Planchon and David A Boothman, Department of Radiation Oncology, CWRU, for providing the DU-145-Bcl-2 cells. We thank Catherine Hollerbach for constructing the plasmid encoding the cytoplasmic version of Bcl-xL. This research was supported by US Public Health Service Grants P01 CA48735, R01 CA83917, and P30 CA43703 from the National Cancer Institute, DHHS, and Grant MT12517 from the Canadian Institutes of Health Research. DWA holds the Canada Research Chair in Membrane Biogenesis.

Author information

Authors and Affiliations

  1. Department of Radiation Oncology, Case Western Reserve University School of Medicine, Cleveland, OH, USA

    Liang-yan Xue, Song-mao Chiu & Nancy L Oleinick

  2. CWRU/Ireland Comprehensive Cancer Center, Case Western Reserve University School of Medicine, Cleveland, OH, USA

    Nancy L Oleinick

  3. Department of Biochemistry, McMaster University, Hamilton, ONT L8N 3Z5, Canada

    Aline Fiebig & David W Andrews

Authors
  1. Liang-yan Xue
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Song-mao Chiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Aline Fiebig
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. David W Andrews
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Nancy L Oleinick
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Nancy L Oleinick.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Xue, Ly., Chiu, Sm., Fiebig, A. et al. Photodamage to multiple Bcl-xL isoforms by photodynamic therapy with the phthalocyanine photosensitizer Pc 4. Oncogene 22, 9197–9204 (2003). https://doi.org/10.1038/sj.onc.1207019

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1207019

Keywords

This article is cited by

Search

Advanced search

Quick links