Reorganization of cytoskeleton induced by 5-aminolevulinic acid-mediated photodynamic therapy and its correlation with mitochondrial dysfunction

Jui-Chang Tsai, Chia-Lun Wu, Hsiung-Fei Chien, Chin-Tin Chen

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Background and Objectives: This study investigated the early cellular events which occurred after mitochondrial photodamage induced by 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT). Study Design/Materials and Methods: Subcellular localization of protoporphyrin IX (PpIX) in NIH3T3 cells was studied by confocal microscopy. Mitochondrial damage was assessed by measuring mitochondrial transmembrane potential and ATP contents, and confirmed by characteristic appearance on transmission electron microscopy. Cellular adhesion was measured by the level of resistance to trypsinization. Cytoskeletal studies were performed by fluorescent staining of cytoskeletal components. Results: Following ALA-PDT, mitochondrial damage was found in NIH3T3 cells as judged by the decrease of membrane potential and ATP contents. Mitochondrial photodamage was futher confirmed by electron microscopy. Resistance to trypsinization after ALA-PDT was shown to be light dose-dependent. The increase of cellular adhesion after ALA-PDT was correlated with mitochondrial photodamage and reorganization of cytoskeletal components in NIH3T3 cells. Conclusions: This study has demonstrated that mitochondrial dysfunctions induced by ALA-PDT results in alterations of cellular morphology and cellular adhesion. © 2005 Wiley-Liss, Inc.
Original languageEnglish
Pages (from-to)398-408
Number of pages11
JournalLasers in Surgery and Medicine
Volume36
Issue number5
DOIs
Publication statusPublished - 2005
Externally publishedYes

Fingerprint

Aminolevulinic Acid
Photochemotherapy
Cytoskeleton
Membrane Potentials
Adenosine Triphosphate
Transmission Electron Microscopy
Confocal Microscopy
Electron Microscopy
Staining and Labeling
Light

Keywords

  • Adhesion
  • Mitochondria
  • Photodynamic therapy
  • Protoporphyrin IX
  • adenosine triphosphate
  • aminolevulinic acid
  • fluorescent dye
  • protoporphyrin
  • animal cell
  • article
  • cell adhesion
  • cell damage
  • cell line
  • confocal microscopy
  • cytoskeleton
  • drug effect
  • membrane potential
  • mitochondrion
  • nonhuman
  • photodynamic therapy
  • priority journal
  • transmission electron microscopy
  • Acid Phosphatase
  • Adenosine Triphosphate
  • Aminolevulinic Acid
  • Animals
  • Cell Adhesion
  • Cytoskeleton
  • Membrane Potentials
  • Mice
  • Microscopy, Electron, Transmission
  • Microscopy, Video
  • NIH 3T3 Cells
  • Photochemotherapy
  • Photosensitizing Agents
  • Protoporphyrins

Cite this

Reorganization of cytoskeleton induced by 5-aminolevulinic acid-mediated photodynamic therapy and its correlation with mitochondrial dysfunction. / Tsai, Jui-Chang; Wu, Chia-Lun; Chien, Hsiung-Fei; Chen, Chin-Tin.

In: Lasers in Surgery and Medicine, Vol. 36, No. 5, 2005, p. 398-408.

Research output: Contribution to journalArticle

@article{ec5d1e9369a847608cbb9f2acc3a5a13,
title = "Reorganization of cytoskeleton induced by 5-aminolevulinic acid-mediated photodynamic therapy and its correlation with mitochondrial dysfunction",
abstract = "Background and Objectives: This study investigated the early cellular events which occurred after mitochondrial photodamage induced by 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT). Study Design/Materials and Methods: Subcellular localization of protoporphyrin IX (PpIX) in NIH3T3 cells was studied by confocal microscopy. Mitochondrial damage was assessed by measuring mitochondrial transmembrane potential and ATP contents, and confirmed by characteristic appearance on transmission electron microscopy. Cellular adhesion was measured by the level of resistance to trypsinization. Cytoskeletal studies were performed by fluorescent staining of cytoskeletal components. Results: Following ALA-PDT, mitochondrial damage was found in NIH3T3 cells as judged by the decrease of membrane potential and ATP contents. Mitochondrial photodamage was futher confirmed by electron microscopy. Resistance to trypsinization after ALA-PDT was shown to be light dose-dependent. The increase of cellular adhesion after ALA-PDT was correlated with mitochondrial photodamage and reorganization of cytoskeletal components in NIH3T3 cells. Conclusions: This study has demonstrated that mitochondrial dysfunctions induced by ALA-PDT results in alterations of cellular morphology and cellular adhesion. {\circledC} 2005 Wiley-Liss, Inc.",
keywords = "Adhesion, Mitochondria, Photodynamic therapy, Protoporphyrin IX, adenosine triphosphate, aminolevulinic acid, fluorescent dye, protoporphyrin, animal cell, article, cell adhesion, cell damage, cell line, confocal microscopy, cytoskeleton, drug effect, membrane potential, mitochondrion, nonhuman, photodynamic therapy, priority journal, transmission electron microscopy, Acid Phosphatase, Adenosine Triphosphate, Aminolevulinic Acid, Animals, Cell Adhesion, Cytoskeleton, Membrane Potentials, Mice, Microscopy, Electron, Transmission, Microscopy, Video, NIH 3T3 Cells, Photochemotherapy, Photosensitizing Agents, Protoporphyrins",
author = "Jui-Chang Tsai and Chia-Lun Wu and Hsiung-Fei Chien and Chin-Tin Chen",
note = "被引用次數:36 Export Date: 16 March 2016 CODEN: LSMED 通訊地址: Chen, C.-T.; Center for Optoelectronic Biomedicine, College of Medicine, National Taiwan University, No. 1, Jen-Ai Road, Section 1st, Taipei 100, Taiwan; 電子郵件: ctchen@ha.mc.ntu.edu.tw 化學物質/CAS: adenosine triphosphate, 15237-44-2, 56-65-5, 987-65-5; aminolevulinic acid, 106-60-5; protoporphyrin, 553-12-8; Acid Phosphatase, EC 3.1.3.2; Adenosine Triphosphate, 56-65-5; Aminolevulinic Acid, 106-60-5; Photosensitizing Agents; protoporphyrin IX, 553-12-8; Protoporphyrins 參考文獻: Dougherty, T.J., Gomer, C.J., Henderson, B.W., Jori, G., Kessel, D., Korbelik, M., Moan, J., Peng, Q., Photodynamic therapy (1998) J Natl Cancer Inst, 90, pp. 889-905; Dolmans, D.E., Fukumura, D., Jain, R.K., Photodynamic therapy for cancer (2003) Nature Rev Cancer, 3, pp. 380-387; Schuitmaker, J.J., Baas, P., Van Leengoed, H.L., Van Der Meulen, F.W., Star, W.M., Van Zandwijk, N., Photodynamic therapy: A promising new modality for the treatment of cancer (1996) J Photochem Photobiol B, 34, pp. 3-12; Henderson, B.W., Dougherty, T.J., How does photodynamic therapy work? (1992) Photochem Photobiol, 55, pp. 145-157; Fisher, A.M., Murphree, A.L., Gomer, C.J., Clinical and preclinical photodynamic therapy (1995) Lasers Surg Med, 17, pp. 2-31; Oleinick, N.L., Evans, H.H., The photobiology of photodynamic therapy: Cellular targets and mechanisms (1998) Radiat Res, 150 (SUPPL. 5), pp. S146-S156; Moor, A.C., Signaling pathways in cell death and survival after photodynamic therapy (2000) J Photochem Photobiol B, 57 (1), pp. 1-13; Gomer, C.J., Rucker, N., Ferrario, A., Wong, S., Properties and applications of photodynamic therapy (1989) Radiat Res, 120, pp. 1-18; Boyle, R.W., Dolphin, D., Structure and biodistribution relationships of photodynamic sensitizers (1996) Photochem Photobiol, 64, pp. 469-485; Gardner, L.C., Smith, S.J., Cox, T.M., Biosynthesis of delta-aminolevulinic acid and the regulation of heme formation by immature erythroid cells in man (1991) J Biol Chem, 266, pp. 22010-22018; Dailey, H.A., Smith, A., Differential interaction of porphyrins used in photoradiation therapy with ferrochelatase (1984) Biochem J, 223, pp. 441-445; Iinuma, S., Farshi, S.S., Ortel, B., Hasan, T., A mechanistic study of cellular photodestruction with 5-aminolaevulinic acid-induced porphyrin (1994) Br J Cancer, 70, pp. 21-28; Gibson, S.L., Nguyen, M.L., Havens, J.J., Barbarin, A., Hilf, R., Relationship of delta-aminolevulinic acid-induced protoporphyrin IX levels to mitochondrial content in neoplastic cells in vitro (1999) Biochem Biophys Res Commun, 265, pp. 315-321; Peng, Q., Warloe, T., Berg, K., Moan, J., Kongshaug, M., Giercksky, K.E., Nesland, J.M., 5-Aminolevulinic acid-based photodynamic therapy: Clinical research and future challenges (1997) Cancer, 79, pp. 2282-2308; Morgan, J., Oseroff, A.B., Mitochondria-based photodynamic anti-cancer therapy (2001) Adv Drug Deliv Rev, 49, pp. 71-86; Almeida, R.D., Manadas, B.J., Carvalho, A.P., Duarte, C.B., Intracellular signaling mechanisms in photodynamic therapy (2004) Biochim Biophys Acta (BBA)-Rev Cancer, 1704, p. 59; Korbelik, M., Dougherty, G.J., Photodynamic therapy-mediated immune response against subcutaneous mouse tumors (1999) Cancer Res, 59, pp. 1941-1946; Gollnick, S.O., Vaughan, L., Henderson, B.W., Generation of effective antitumor vaccines using photodynamic therapy (2002) Cancer Res, 62, pp. 1604-1608; Oleinick, N.L., Morris, R.L., Belichenko, I., The role of apoptosis in response to photodynamic therapy: What, where, why, and how (2002) Photochem Photobiol Sci, 1, pp. 1-21; Tsai, J.C., Chiang, C.P., Chen, H.M., Huang, S.B., Wang, C.W., Lee, M.I., Hsu, Y.C., Tsai, T., Photodynamic Therapy of oral dysplasia with topical 5-aminolevulinic acid and light-emitting diode array (2004) Lasers Surg Med, 34, pp. 18-24; Mosmann, T., Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays (1983) J Immunol Methods, 65, pp. 55-63; Yang, T.T., Sinai, P., Kain, S.R., An acid phosphatase assay for quantifying the growth of adherent and nonadherent cells (1996) Anal Biochem, 241, pp. 103-108; Tsai, T., Hong, L.-M., Lou, P.-J., Ling, I.-F., Chen, C.T., Effect of 5-aminolevulinic acid-mediated photodynamic therapy on MCF-7 and MCF-7/ADR cells (2004) Lasers Surg Med, 34, pp. 62-72; Moan, J., Berg, K., The photodegradation of porphyrins in cells can be used to estimate the lifetime of singlet oxygen (1991) Photochem Photobiol, 53, pp. 549-553; Krammer, B., Uberriegler, K., In-vitro investigation of ALA-induced protoporphyrin IX (1996) J Photochem Photobiol B, 36, pp. 121-126; Kuzelova, K., Grebenova, D., Pluskalova, M., Marinov, I., Hrkal, Z., Early apoptotic features of K562 cell death induced by 5-aminolaevulinic acid-based photodynamic therapy (2004) J Photochem Photobiol B, 73, pp. 67-78; Foultier, M.T., Vonarx-Coinsmann, V., Cordel, S., Combre, A., Patrice, T., Modulation of colonic cancer cell adhesiveness by haematoporphyrin derivative photodynamic therapy (1994) J Photochem Photobiol B, 23, pp. 9-17; Denstman, S.C., Dillehay, L.E., Williams, J.R., Enhanced susceptibility to HPD-sensitized phototoxicity and correlated resistance to trypsin detachment in SV40 transformed IMR-90 cells (1986) Photochem Photobiol, 43, pp. 145-147; Hunting, D.J., Gowans, B.J., Brasseur, N., Van Lier, J.E., DNA damage and repair following treatment of V-79 cells with sulfonated phthalocyanines (1987) Photochem Photobiol, 45, pp. 769-773; Ball, D.J., Mayhew, S., Vernon, D.I., Griffin, M., Brown, S.B., Decreased efficiency of trypsinization of cells following photodynamic therapy: Evaluation of a role for tissue transglutaminase (2001) Photochem Photobiol, 73, pp. 47-53; Teiten, M.H., Marchai, S., D'Hallewin, M.A., Guillemin, F., Bezdetnaya, L., Primary photodamage sites and mitochondrial events after Foscan photosensitization of MCF-7 human breast cancer cells (2003) Photochem Photobiol, 78, pp. 9-14; Ball, D.J., Mayhew, S., Wood, S.R., Griffiths, J., Vernon, D.I., Brown, S.B., A comparative study of the cellular uptake and photodynamic efficacy of three novel zinc phthalocyanines of differing charge (1999) Photochem Photobiol, 69, pp. 390-396; Ben-Hur, E., Rosenthal, I., Leznoff, C.C., Recovery of Chinese hamster cells following photosensitization by zinc tetrahydroxyphthalocyanine (1988) J Photochem Photobiol B, 2, pp. 243-252; Christensen, T., Moan, J., Smedshammer, L., Western, A., Rimington, C., Influence of hematoporphyrin derivative (HpD) and light on the attachment of cells to the substratum (1985) Photobiochem Photobiophys, 10, pp. 53-59; Morgan, J., Potter, W.R., Oseroff, A.R., Comparison of photodynamic targets in a carcinoma cell line and its mitochondrial DNA-deficient derivative (2000) Photochem Photobiol, 71, pp. 747-757; Seagrave, J.C., Burchiel, S.W., Interactions between benzo[a]-pyrene and UVA light affecting ATP levels, cytoskeletal organization, and resistance to trypsinization (2000) Toxicol Lett, 117, pp. 11-23; Janmey, P.A., The cytoskeleton and cell signaling: Component localization and mechanical coupling (1998) Physiol Rev, 78, pp. 763-781",
year = "2005",
doi = "10.1002/lsm.20179",
language = "English",
volume = "36",
pages = "398--408",
journal = "Lasers in Surgery and Medicine",
issn = "0196-8092",
publisher = "Wiley-Liss Inc.",
number = "5",

}

TY - JOUR

T1 - Reorganization of cytoskeleton induced by 5-aminolevulinic acid-mediated photodynamic therapy and its correlation with mitochondrial dysfunction

AU - Tsai, Jui-Chang

AU - Wu, Chia-Lun

AU - Chien, Hsiung-Fei

AU - Chen, Chin-Tin

N1 - 被引用次數:36 Export Date: 16 March 2016 CODEN: LSMED 通訊地址: Chen, C.-T.; Center for Optoelectronic Biomedicine, College of Medicine, National Taiwan University, No. 1, Jen-Ai Road, Section 1st, Taipei 100, Taiwan; 電子郵件: ctchen@ha.mc.ntu.edu.tw 化學物質/CAS: adenosine triphosphate, 15237-44-2, 56-65-5, 987-65-5; aminolevulinic acid, 106-60-5; protoporphyrin, 553-12-8; Acid Phosphatase, EC 3.1.3.2; Adenosine Triphosphate, 56-65-5; Aminolevulinic Acid, 106-60-5; Photosensitizing Agents; protoporphyrin IX, 553-12-8; Protoporphyrins 參考文獻: Dougherty, T.J., Gomer, C.J., Henderson, B.W., Jori, G., Kessel, D., Korbelik, M., Moan, J., Peng, Q., Photodynamic therapy (1998) J Natl Cancer Inst, 90, pp. 889-905; Dolmans, D.E., Fukumura, D., Jain, R.K., Photodynamic therapy for cancer (2003) Nature Rev Cancer, 3, pp. 380-387; Schuitmaker, J.J., Baas, P., Van Leengoed, H.L., Van Der Meulen, F.W., Star, W.M., Van Zandwijk, N., Photodynamic therapy: A promising new modality for the treatment of cancer (1996) J Photochem Photobiol B, 34, pp. 3-12; Henderson, B.W., Dougherty, T.J., How does photodynamic therapy work? (1992) Photochem Photobiol, 55, pp. 145-157; Fisher, A.M., Murphree, A.L., Gomer, C.J., Clinical and preclinical photodynamic therapy (1995) Lasers Surg Med, 17, pp. 2-31; Oleinick, N.L., Evans, H.H., The photobiology of photodynamic therapy: Cellular targets and mechanisms (1998) Radiat Res, 150 (SUPPL. 5), pp. S146-S156; Moor, A.C., Signaling pathways in cell death and survival after photodynamic therapy (2000) J Photochem Photobiol B, 57 (1), pp. 1-13; Gomer, C.J., Rucker, N., Ferrario, A., Wong, S., Properties and applications of photodynamic therapy (1989) Radiat Res, 120, pp. 1-18; Boyle, R.W., Dolphin, D., Structure and biodistribution relationships of photodynamic sensitizers (1996) Photochem Photobiol, 64, pp. 469-485; Gardner, L.C., Smith, S.J., Cox, T.M., Biosynthesis of delta-aminolevulinic acid and the regulation of heme formation by immature erythroid cells in man (1991) J Biol Chem, 266, pp. 22010-22018; Dailey, H.A., Smith, A., Differential interaction of porphyrins used in photoradiation therapy with ferrochelatase (1984) Biochem J, 223, pp. 441-445; Iinuma, S., Farshi, S.S., Ortel, B., Hasan, T., A mechanistic study of cellular photodestruction with 5-aminolaevulinic acid-induced porphyrin (1994) Br J Cancer, 70, pp. 21-28; Gibson, S.L., Nguyen, M.L., Havens, J.J., Barbarin, A., Hilf, R., Relationship of delta-aminolevulinic acid-induced protoporphyrin IX levels to mitochondrial content in neoplastic cells in vitro (1999) Biochem Biophys Res Commun, 265, pp. 315-321; Peng, Q., Warloe, T., Berg, K., Moan, J., Kongshaug, M., Giercksky, K.E., Nesland, J.M., 5-Aminolevulinic acid-based photodynamic therapy: Clinical research and future challenges (1997) Cancer, 79, pp. 2282-2308; Morgan, J., Oseroff, A.B., Mitochondria-based photodynamic anti-cancer therapy (2001) Adv Drug Deliv Rev, 49, pp. 71-86; Almeida, R.D., Manadas, B.J., Carvalho, A.P., Duarte, C.B., Intracellular signaling mechanisms in photodynamic therapy (2004) Biochim Biophys Acta (BBA)-Rev Cancer, 1704, p. 59; Korbelik, M., Dougherty, G.J., Photodynamic therapy-mediated immune response against subcutaneous mouse tumors (1999) Cancer Res, 59, pp. 1941-1946; Gollnick, S.O., Vaughan, L., Henderson, B.W., Generation of effective antitumor vaccines using photodynamic therapy (2002) Cancer Res, 62, pp. 1604-1608; Oleinick, N.L., Morris, R.L., Belichenko, I., The role of apoptosis in response to photodynamic therapy: What, where, why, and how (2002) Photochem Photobiol Sci, 1, pp. 1-21; Tsai, J.C., Chiang, C.P., Chen, H.M., Huang, S.B., Wang, C.W., Lee, M.I., Hsu, Y.C., Tsai, T., Photodynamic Therapy of oral dysplasia with topical 5-aminolevulinic acid and light-emitting diode array (2004) Lasers Surg Med, 34, pp. 18-24; Mosmann, T., Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays (1983) J Immunol Methods, 65, pp. 55-63; Yang, T.T., Sinai, P., Kain, S.R., An acid phosphatase assay for quantifying the growth of adherent and nonadherent cells (1996) Anal Biochem, 241, pp. 103-108; Tsai, T., Hong, L.-M., Lou, P.-J., Ling, I.-F., Chen, C.T., Effect of 5-aminolevulinic acid-mediated photodynamic therapy on MCF-7 and MCF-7/ADR cells (2004) Lasers Surg Med, 34, pp. 62-72; Moan, J., Berg, K., The photodegradation of porphyrins in cells can be used to estimate the lifetime of singlet oxygen (1991) Photochem Photobiol, 53, pp. 549-553; Krammer, B., Uberriegler, K., In-vitro investigation of ALA-induced protoporphyrin IX (1996) J Photochem Photobiol B, 36, pp. 121-126; Kuzelova, K., Grebenova, D., Pluskalova, M., Marinov, I., Hrkal, Z., Early apoptotic features of K562 cell death induced by 5-aminolaevulinic acid-based photodynamic therapy (2004) J Photochem Photobiol B, 73, pp. 67-78; Foultier, M.T., Vonarx-Coinsmann, V., Cordel, S., Combre, A., Patrice, T., Modulation of colonic cancer cell adhesiveness by haematoporphyrin derivative photodynamic therapy (1994) J Photochem Photobiol B, 23, pp. 9-17; Denstman, S.C., Dillehay, L.E., Williams, J.R., Enhanced susceptibility to HPD-sensitized phototoxicity and correlated resistance to trypsin detachment in SV40 transformed IMR-90 cells (1986) Photochem Photobiol, 43, pp. 145-147; Hunting, D.J., Gowans, B.J., Brasseur, N., Van Lier, J.E., DNA damage and repair following treatment of V-79 cells with sulfonated phthalocyanines (1987) Photochem Photobiol, 45, pp. 769-773; Ball, D.J., Mayhew, S., Vernon, D.I., Griffin, M., Brown, S.B., Decreased efficiency of trypsinization of cells following photodynamic therapy: Evaluation of a role for tissue transglutaminase (2001) Photochem Photobiol, 73, pp. 47-53; Teiten, M.H., Marchai, S., D'Hallewin, M.A., Guillemin, F., Bezdetnaya, L., Primary photodamage sites and mitochondrial events after Foscan photosensitization of MCF-7 human breast cancer cells (2003) Photochem Photobiol, 78, pp. 9-14; Ball, D.J., Mayhew, S., Wood, S.R., Griffiths, J., Vernon, D.I., Brown, S.B., A comparative study of the cellular uptake and photodynamic efficacy of three novel zinc phthalocyanines of differing charge (1999) Photochem Photobiol, 69, pp. 390-396; Ben-Hur, E., Rosenthal, I., Leznoff, C.C., Recovery of Chinese hamster cells following photosensitization by zinc tetrahydroxyphthalocyanine (1988) J Photochem Photobiol B, 2, pp. 243-252; Christensen, T., Moan, J., Smedshammer, L., Western, A., Rimington, C., Influence of hematoporphyrin derivative (HpD) and light on the attachment of cells to the substratum (1985) Photobiochem Photobiophys, 10, pp. 53-59; Morgan, J., Potter, W.R., Oseroff, A.R., Comparison of photodynamic targets in a carcinoma cell line and its mitochondrial DNA-deficient derivative (2000) Photochem Photobiol, 71, pp. 747-757; Seagrave, J.C., Burchiel, S.W., Interactions between benzo[a]-pyrene and UVA light affecting ATP levels, cytoskeletal organization, and resistance to trypsinization (2000) Toxicol Lett, 117, pp. 11-23; Janmey, P.A., The cytoskeleton and cell signaling: Component localization and mechanical coupling (1998) Physiol Rev, 78, pp. 763-781

PY - 2005

Y1 - 2005

N2 - Background and Objectives: This study investigated the early cellular events which occurred after mitochondrial photodamage induced by 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT). Study Design/Materials and Methods: Subcellular localization of protoporphyrin IX (PpIX) in NIH3T3 cells was studied by confocal microscopy. Mitochondrial damage was assessed by measuring mitochondrial transmembrane potential and ATP contents, and confirmed by characteristic appearance on transmission electron microscopy. Cellular adhesion was measured by the level of resistance to trypsinization. Cytoskeletal studies were performed by fluorescent staining of cytoskeletal components. Results: Following ALA-PDT, mitochondrial damage was found in NIH3T3 cells as judged by the decrease of membrane potential and ATP contents. Mitochondrial photodamage was futher confirmed by electron microscopy. Resistance to trypsinization after ALA-PDT was shown to be light dose-dependent. The increase of cellular adhesion after ALA-PDT was correlated with mitochondrial photodamage and reorganization of cytoskeletal components in NIH3T3 cells. Conclusions: This study has demonstrated that mitochondrial dysfunctions induced by ALA-PDT results in alterations of cellular morphology and cellular adhesion. © 2005 Wiley-Liss, Inc.

AB - Background and Objectives: This study investigated the early cellular events which occurred after mitochondrial photodamage induced by 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT). Study Design/Materials and Methods: Subcellular localization of protoporphyrin IX (PpIX) in NIH3T3 cells was studied by confocal microscopy. Mitochondrial damage was assessed by measuring mitochondrial transmembrane potential and ATP contents, and confirmed by characteristic appearance on transmission electron microscopy. Cellular adhesion was measured by the level of resistance to trypsinization. Cytoskeletal studies were performed by fluorescent staining of cytoskeletal components. Results: Following ALA-PDT, mitochondrial damage was found in NIH3T3 cells as judged by the decrease of membrane potential and ATP contents. Mitochondrial photodamage was futher confirmed by electron microscopy. Resistance to trypsinization after ALA-PDT was shown to be light dose-dependent. The increase of cellular adhesion after ALA-PDT was correlated with mitochondrial photodamage and reorganization of cytoskeletal components in NIH3T3 cells. Conclusions: This study has demonstrated that mitochondrial dysfunctions induced by ALA-PDT results in alterations of cellular morphology and cellular adhesion. © 2005 Wiley-Liss, Inc.

KW - Adhesion

KW - Mitochondria

KW - Photodynamic therapy

KW - Protoporphyrin IX

KW - adenosine triphosphate

KW - aminolevulinic acid

KW - fluorescent dye

KW - protoporphyrin

KW - animal cell

KW - article

KW - cell adhesion

KW - cell damage

KW - cell line

KW - confocal microscopy

KW - cytoskeleton

KW - drug effect

KW - membrane potential

KW - mitochondrion

KW - nonhuman

KW - photodynamic therapy

KW - priority journal

KW - transmission electron microscopy

KW - Acid Phosphatase

KW - Adenosine Triphosphate

KW - Aminolevulinic Acid

KW - Animals

KW - Cell Adhesion

KW - Cytoskeleton

KW - Membrane Potentials

KW - Mice

KW - Microscopy, Electron, Transmission

KW - Microscopy, Video

KW - NIH 3T3 Cells

KW - Photochemotherapy

KW - Photosensitizing Agents

KW - Protoporphyrins

U2 - 10.1002/lsm.20179

DO - 10.1002/lsm.20179

M3 - Article

VL - 36

SP - 398

EP - 408

JO - Lasers in Surgery and Medicine

JF - Lasers in Surgery and Medicine

SN - 0196-8092

IS - 5

ER -