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Effects of BAPTA-AM, Forskolin, DSF and Z.VAD.fmk on PDT-induced apoptosis and m-THPC phototoxicity on B16 cells

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Abstract

As many types of cells exposed to photodynamic therapy (PDT) appear to undergo apoptosis, various apoptosis inhibitors have already been used in studies of PDT-induced apoptosis. Although these inhibitors decrease apoptosis, their real effect on the phototoxic efficacy of photosensitisers is unclear. The good phototoxicity of m-THPC was confirmed on murine melanoma B16-A45 cells. Toxicity and phototoxicity studies were then carried out using four apoptosis inhibitors: BAPTA-AM, Forskolin, DSF, and Z.VAD.fmk. Although all inhibitors tested blocked PDT-induced apoptosis, none produced a significant modification of the phototoxic effect of m-THPC on B16 cells. It has been suggested that apoptosis and necrosis share common initiation pathways and that the final outcome is determined by the presence of an active caspase. This implies that apoptosis inhibition reorients cells to necrosis, i.e. those cells sufficiently damaged by PDT appear to be killed, regardless of the mechanism involved.

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References

  1. Weishaupt KR, Gomer CJ, Dougherty TJ Identification of singlet oxygen as the cytotoxic agent in photoinactivation of murine tumors. Cancer Res 1976; 36: 2326–2329.

    Google Scholar 

  2. Mitchell J, McPherson S, Degraff W, Gamson J, Zabell A, Russo A Oxygen dependence of hematoporphyrin derivativeinduced photoinactivation of Chinese hamster cells. Cancer Res 1985; 45: 2008–2011.

    Google Scholar 

  3. Kerr JFR, Wyllie AH Apoptosis: A basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 1972; 26: 239–257.

    Google Scholar 

  4. Wyllie AH The biology of cell death in tumors. Anticancer Res 1985; 5: 131–136.

    Google Scholar 

  5. Vaux DL, Korsmeyer SJ Cell death in development. Cell 1999; 96: 245–254.

    Google Scholar 

  6. Waring P, Kos FJ, Mullbacher A Apoptosis or programmed cell death. Med Res Rev 1991; 11: 219–236.

    Google Scholar 

  7. Sheridan JW, Bishop CJ, Simmons RJ Biophysical and morphological correlates of kinetic changes and death in a starved human melanoma cell line. J Cell Sci 1981; 49: 119–137.

    Google Scholar 

  8. Manson MD Cellular aspects of photodynamic therapy for cancer. Rev Contemp Pharmacother 1999; 10: 25–37.

    Google Scholar 

  9. Agarwal ML, Clay ME, Harvey EJ, Evans HH, Antunez AR, Oleinick NL Photodynamic therapy induces rapid cell death by apoptosis in L5178Y mouse lymphoma cells. Cancer Res 1991; 51: 5993–5996.

    Google Scholar 

  10. He XY, Sikes RA, Thomsen S, Chung LW, Jacques SL Photodynamic therapy with Photofrin II induces programmed cell death in carcinoma cell lines. Photochem Photobiol 1994; 59: 468–473.

    Google Scholar 

  11. Zaidi SI, Oleinick NL, Zaim MT, Mukhtar H Apoptosis during photodynamic therapy-induced ablation of RIF-1 tumors in C3H mice: Electron microscopic, histopathologic and biochemical evidence. Photochem Photobiol 1993; 58: 771–776.

    Google Scholar 

  12. Webber J, Luo Y, Crilly R, Fromm D, Kessel D An apoptotic response to photodynamic therapy with endogenous protoporphyrin in vivo. J Photochem Photobiol B 1996; 35: 209–211.

    Google Scholar 

  13. Inanami O, Yoshito A, Takahashi K, Hiraoka W, Kuwabara M Effects of BAPTA-AM and Forskolin on apoptosis and cytochrome c release in photosensitized Chinese hamster V79 cells. Photochem Photobiol 1999; 70: 650–655.

    Google Scholar 

  14. Iwashita K, Kobori M, Yamaki K, Tsushida T Flavonoids inhibit cell growth and induce apoptosis in B16 melanoma 4A5 cells. Biosci Biotechnol Biochem 2000; 64: 1813–1820.

    Google Scholar 

  15. Sredni B, Longo DL, Catane R, Shani A, Albeck M, Kalechman Y Synergistic antitumoral effect of the immunomodulator AS101+paclitaxel (Taxol). Association with ras-dependent signal transduction pathways. Exp Hematol 2000; 28: 1496.

    Google Scholar 

  16. Vonarx-Coinsmann V, Foultier MT, Cempel N, Morlet L, Combre A, Patrice T In vitro and in vivo photodynamic effects of a new photosensitizer: Tetra(m-hydroxyphenyl)chlorin. Lasers Med Sci 1994; 9: 173–181.

    Google Scholar 

  17. Dilkes MG, Dejode ML, Rowntree-Taylor A m-THPC photodynamic therapy for head and neck cancer. Lasers Med Sci 1997; 11: 23–30.

    Google Scholar 

  18. Chen JY, Mak NK, Wen JM, et al Acomparison of the photodynamic effects of temoporfin (mTHPC) and MC540 on leukemia cells: Efficacy and apoptosis. Photochem Photobiol 1998; 68: 545–554.

    Google Scholar 

  19. Lilge L, Portnoy M, Wilson BC Apoptosis induced in vivo by photodynamic therapy in normal brain and intracranial tumour tissue. Br J Cancer 2000; 83: 1110–1117.

    Google Scholar 

  20. Lemaire C, Andreau K, Souvannavong V, Adam A Inhibition of caspase activity induces a switch from apoptosis to necrosis. FEBS Letters 1998; 425: 266–270.

    Google Scholar 

  21. Samali A, Nordgren, Zhivotovsky B, Peterson E, Orrenius S A comparative study of apoptosis and necrosis in HepG2 cells: Oxidant-induced caspase inactivation leads to necrosis. Biochem Biophys Res Com 1999; 255: 6–11.

    Google Scholar 

  22. Burkitt MJ, Bishop HS, Milne L, et al Dithiocarbamate toxicity toward thymocytes involves their copper-catalyzed conversion to thiuram disulfides, which oxidize glutathione in a redox cycle without the release of reactive oxygen species. Arch Biochem Biophys 1998; 353: 73–84.

    Google Scholar 

  23. Nobel CSI, Burgess DH, Zhivotovsky B, Burkitt MJ, OrreniuS S, Slater AFG Mechanism of dithiocarbamate inhibition of apoptosis: Thiol oxidation by dithiocarbamate disulfides directly inhibits processing of the caspase-3 proenzyme. Chem Res Toxicol 1997; 10: 636–643.

    Google Scholar 

  24. Haddad R, Blumenfeld A, Siegal A, et al In vitro and in vivo effects of photodynamic therapy on murine malignant melanoma. Ann Surg Oncol 1998; 5: 241–247.

    Google Scholar 

  25. Mosmann T Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assay. J Immunol Methods 1983; 65: 55–63.

    Google Scholar 

  26. Boiadjieva S, Hallberg C, Högström M, Busch C Exclusion of Trypan Blue from microcarriers by endothelial cells: An in vitro barrier function test. Met Lab Invest 1984; 50: 239–246.

    Google Scholar 

  27. Bachor R, Shea CR, Belmonte SJ, Hasan T Free and conjugated chlorin E6 in the photodynamic therapy of human blabber carcinoma cells. J Uro 1991; 146: 1654–1658.

    Google Scholar 

  28. Salgame P, Varadhachary AS, Primiano L, Fincke JE, Muller S, Monestier An ELISA for detection of apoptosis. Nucleic Acids Res 1997; 25: 680–683.

    Google Scholar 

  29. Savary JF, Grosjean P, Monnier P, et al Photodynamic therapy of early squamous cell carcinomas of the esophagus: A review of 31 cases. Endoscopy 1998; 30: 258–265.

    Google Scholar 

  30. Kostron H, Obwegeser A, Jakober R Photodynamic therapy in neurosurgery: A review. J Photochem Photobiol B 1996; 36: 157–168.

    Google Scholar 

  31. Ell C, Gossner L, May A, et al Photodynamic ablation of early cancers of the stomach by means of m-THPC and laser irradiation: Preliminary clinical experience. Gut 1998; 43: 345–349.

  32. Laukka MA, Wang KK, Bonner JA Apoptosis occurs in lymphoma cells but not in hepatoma cells following ionizing radiation and photodynamic therapy. Digest Dis Sci 1994; 39: 2467–2475.

    Google Scholar 

  33. Kessel D, Luo Y Delayed oxidative photodamage induced by photodynamic therapy. Photochem Photobiol 1996; 64: 601–604.

    Google Scholar 

  34. Ahmad N, Feyes DK, Agarwal R, Mukhtar H Photodynamic therapy results in induction ofWAF1/CIPI/P21 leading to cell arrest and apoptosis. Proc Natl Acad Sci USA 1998; 95: 6977–6982.

    Google Scholar 

  35. Guilian N, Heller R, Catlett-Falcone R, et al Gene therapy with dominant-negative Stat3 suppresses growth of the murine melanoma B16 tumor in vivo. Cancer Res 1999; 5: 5059–5063.

    Google Scholar 

  36. Tsai JC, Hsiao YY, Teng LJ, Chen CT, Kao MC Comparative study on the ALA photodynamic effects of human glioma and meningioma cells. Lasers Surg Med 1999; 24: 296–305.

    Google Scholar 

  37. Sheyhedin I, Aizawa K, Araake M, Kumasaka H, Okunaka T, Kato H The effects of serum on cellular uptake and phototoxicity of mono-L-aspartyl chlorin e6 (NPe6) in vitro. Photochem Photobiol 1998; 68: 110–114.

    Google Scholar 

  38. Tajiri H, Hayakawa A, Matsumoto Y, Yokoyama I, Yoshida S Changes in intracellular Ca2+ concentrations related to PDTinduced apoptosis in photosensitized human cancer cells. Cancer Lett 1998; 128: 205–210.

    Google Scholar 

  39. Liu X, Kim CN, Jemmerson R, Wang X Induction of apoptotic program in cell-free extracts: Requirement for dATP and cytochrome c. Cell 1996; 86: 147–157.

    Google Scholar 

  40. Burgoyne LA, Hewish DR, Mobbs J Mammalian chromatin substructure studies with the calcium-magnesium endonuclease and two-dimensional polyacrylamide-gel electrophoresis. Biochem J 1974; 143: 67–72.

    Google Scholar 

  41. Sumantran VN, Ealovega MW, Nunez G, Clarke MF, Wicha MS Overexpression of Bcl-xs sensitizes MCF-7 cells to chemotherapy-induced apoptosis. Cancer Res 1995; 55: 2507–2510.

    Google Scholar 

  42. Mandal M, Wu X, Kumar R Bcl-2 deregulation leads to inhibition of sodium butyrate-induced apoptosis in human colorectal carcinoma cells. Carcinogenesis 1997; 18: 229–232.

    Google Scholar 

  43. Lennon SV, Kilfeather SA, Cotter TG The role of calcium in the induction of apoptosis in human tumour cell lines. Biochem Soc Trans 1992; 20: 77S.

    Google Scholar 

  44. Rodriguez-Tarduchy G, Malde P, Lopez-Rivas A, Collins MKL Inhibition of apoptosis by calcium ionophores in IL-3-dependent bone marrow cells is dependent upon production of IL-4. J Immunol 1992; 148: 1416–1422.

    Google Scholar 

  45. Lennon SV, Martin SJ, Cotter TG Dose-dependent induction of apoptosis in human tumour cell lines by widely diverging stimuli. Cell Prolif 1991; 24: 203–214.

    Google Scholar 

  46. Luo Y, Kessel D Initiation of apoptosis versus necrosis by photodynamic therapy with chloroaluminium phtalocyanine. Photochem Photobiol 1997; 66: 479–483.

    Google Scholar 

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Authors and Affiliations

  1. Département Laser, Hôpital Laënnec, 44093, Nantes, France

    S. Thibaut, L. Bourré, D. Hernot, N. Rousset, Y. Lajat & T. Patrice

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  1. S. Thibaut
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  2. L. Bourré
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  3. D. Hernot
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  4. N. Rousset
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Thibaut, S., Bourré, L., Hernot, D. et al. Effects of BAPTA-AM, Forskolin, DSF and Z.VAD.fmk on PDT-induced apoptosis and m-THPC phototoxicity on B16 cells. Apoptosis 7, 99–106 (2002). https://doi.org/10.1023/A:1014350128251

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