Photodynamic therapy inhibits cell adhesion without altering integrin expression.

Adhesion is a primordial cell function that, among others, regulates inflammation, metastasis, and tissue repair. To understand how these events could be affected by photodynamic therapy (PDT), we studied the effects of PDT on human foreskin fibroblast (HFF) adhesion to bovine collagen type I, human vitronectin or fibronectin. PDT, using benzoporphyrin derivative monoacid ring A (verteporfin) as the photosensitizer, inhibited cell adhesion in a drug dose-dependent manner, with no significant difference among matrices. The drug dose that killed 90% of cells within 20 h post-treatment inhibited HFF adhesion by 55%-68%. However, 45 min following PDT, a time period corresponding to that of the adhesion assay, HFF membrane integrity remained unaltered. In addition, cell surface expression of integrins was not modified for at least 2h following PDT. Western blots of cell lysates, using the anti-phosphotyrosine 4G10 monoclonal antibody, revealed that PDT prevented the adhesion-induced phosphorylation of 110-130 kDa proteins. Immunoblots of cell lysates immunoprecipitated with antibodies to focal adhesion kinase suggested that its phosphorylation was suppressed by PDT. These results demonstrate that PDT inhibits cell adhesion and affects integrin signalling without modifying cell membrane integrity or integrin expression.

[1]  V. Quaranta,et al.  Integrin cytoplasmic domains mediate inside-out signal transduction , 1994, The Journal of cell biology.

[2]  A. Girotti,et al.  Photodynamic action of bilirubin on liposomes and erythrocyte membranes. , 1980, The Journal of biological chemistry.

[3]  R. Bates,et al.  Individual embryonic fibroblasts express multiple beta chains in association with the alpha v integrin subunit. Loss of beta 3 expression with cell confluence. , 1991, The Journal of biological chemistry.

[4]  K. Alitalo,et al.  Overexpressed Csk tyrosine kinase is localized in focal adhesions, causes reorganization of alpha v beta 5 integrin, and interferes with HeLa cell spreading , 1995, Molecular and cellular biology.

[5]  Richard O. Hynes,et al.  Integrins: Versatility, modulation, and signaling in cell adhesion , 1992, Cell.

[6]  A. Girotti,et al.  PHOTODYNAMIC LIPID PEROXIDATION IN BIOLOGICAL SYSTEMS * , 1990, Photochemistry and photobiology.

[7]  R. Petty,et al.  Antibodies to native type I collagen in childhood rheumatic diseases. , 1984, The Journal of rheumatology.

[8]  R. Kramer,et al.  β1 And β3 integrins have different roles in the adhesion and migration of vascular smooth muscle cells on extracellular matrix , 1992 .

[9]  T. Patrice,et al.  Modulation of colonic cancer cell adhesiveness by haematoporphyrin derivative photodynamic therapy. , 1994, Journal of photochemistry and photobiology. B, Biology.

[10]  M. Bretscher,et al.  Circulating integrins: alpha 5 beta 1, alpha 6 beta 4 and Mac‐1, but not alpha 3 beta 1, alpha 4 beta 1 or LFA‐1. , 1992, The EMBO journal.

[11]  P B Cerrito,et al.  The role of microvascular damage in photodynamic therapy: the effect of treatment on vessel constriction, permeability, and leukocyte adhesion. , 1992, Cancer research.

[12]  W. Thorpe,et al.  Dynamics of photoinduced cell plasma membrane injury. , 1995, Biophysical journal.

[13]  M. Jalkanen,et al.  Function of the syndecans--a family of cell surface proteoglycans. , 1994, Journal of cell science.

[14]  T. Dubbelman,et al.  Temperature dependence of photodynamic red cell membrane damage. , 1980, Biochimica et biophysica acta.

[15]  C J Gomer,et al.  Properties and applications of photodynamic therapy. , 1989, Radiation research.

[16]  J. Parsons,et al.  Signal transduction through integrins: A central role for focal adhesion kinase? , 1995, BioEssays : news and reviews in molecular, cellular and developmental biology.

[17]  B. Gumbiner,et al.  Cell Adhesion: The Molecular Basis of Tissue Architecture and Morphogenesis , 1996, Cell.

[18]  F. Maxfield,et al.  Ca2+- and calcineurin-dependent recycling of an integrin to the front of migrating neutrophils , 1995, Nature.

[19]  Hedy E. Larkin,et al.  Phospholipase activation triggers apoptosis in photosensitized mouse lymphoma cells. , 1993, Cancer research.

[20]  D. Hinshaw,et al.  Impairment of integrin-mediated cell-matrix adhesion in oxidant-stressed PC12 cells , 1994, Brain Research.

[21]  Y. Yazaki,et al.  Integrin-mediated Cell Adhesion Promotes Tyrosine Phosphorylation of p130, a Src Homology 3-containing Molecule Having Multiple Src Homology 2-binding Motifs (*) , 1995, The Journal of Biological Chemistry.

[22]  S. Habu,et al.  PUVA suppresses the expression of cell adhesion molecules of lymphocytes , 1995, Experimental dermatology.

[23]  J. Parsons,et al.  Focal adhesion kinase and associated proteins. , 1994, Current opinion in cell biology.

[24]  J. Levy,et al.  STRESS PROTEIN EXPRESSION IN MURINE TUMOR CELLS FOLLOWING PHOTODYNAMIC THERAPY WITH BENZOPORPHYRIN DERIVATIVE , 1993, Photochemistry and photobiology.

[25]  R. G. Statius van Eps,et al.  Differential modulation of vascular endothelial and smooth muscle cell function by photodynamic therapy of extracellular matrix: novel insights into radical-mediated prevention of intimal hyperplasia. , 1996, Journal of vascular surgery.

[26]  D. Sheppard,et al.  Role of the integrin alpha v beta 6 in cell attachment to fibronectin. Heterologous expression of intact and secreted forms of the receptor. , 1994, The Journal of biological chemistry.

[27]  M. Jackson,et al.  Quantitative measurement of alpha 6 beta 1 and alpha 6 beta 4 integrin internalization under cross-linking conditions: a possible role for alpha 6 cytoplasmic domains. , 1994, Journal of cell science.

[28]  D. Cheresh,et al.  Purification and functional characterization of integrin alpha v beta 5. An adhesion receptor for vitronectin. , 1990, The Journal of biological chemistry.

[29]  K. Sugimachi,et al.  K-ras and p53 mutations are an independent unfavourable prognostic indicator in patients with non-small-cell lung cancer. , 1997, British Journal of Cancer.

[30]  E. Ruoslahti,et al.  Tyrosine Phosphorylation of p130Cas and Cortactin Accompanies Integrin-mediated Cell Adhesion to Extracellular Matrix (*) , 1995, The Journal of Biological Chemistry.

[31]  E. Land,et al.  Porphyrin localization and treatment of tumors , 1985, British Journal of Cancer.

[32]  Q. Peng,et al.  Correlation of subcellular and intratumoral photosensitizer localization with ultrastructural features after photodynamic therapy. , 1996, Ultrastructural pathology.

[33]  K. Ariizumi,et al.  HYDROGEN PEROXIDE MEDIATES UV‐INDUCED IMPAIRMENT OF ANTIGEN PRESENTATION IN A MURINE EPIDERMAL‐DERIVED DENDRITIC CELL LINE , 1995, Photochemistry and photobiology.

[34]  J. Levy,et al.  Photodynamic therapy induces caspase-3 activation in HL-60 cells , 1997, Cell Death and Differentiation.

[35]  M L Agarwal,et al.  Photodynamic therapy induces rapid cell death by apoptosis in L5178Y mouse lymphoma cells. , 1991, Cancer research.

[36]  S. Pav,et al.  INACTIVATION OF CELL SURFACE RECEPTORS BY PHEOPHORBIDE a, A GREEN PIGMENT ISOLATED FROM Psychotria acuminata , 1995, Photochemistry and photobiology.

[37]  J. Brugge,et al.  Integrins and signal transduction pathways: the road taken. , 1995, Science.

[38]  T. Dougherty,et al.  HOW DOES PHOTODYNAMIC THERAPY WORK? , 1992, Photochemistry and photobiology.

[39]  Y. Yazaki,et al.  Csk enhances insulin-stimulated dephosphorylation of focal adhesion proteins , 1996, Molecular and cellular biology.

[40]  D. Dolphin,et al.  In vitro EVALUATION OF PHOTOTOXIC PROPERTIES OF FOUR STRUCTURALLY RELATED BENZOPORPHYRIN DERIVATIVES , 1990, Photochemistry and photobiology.

[41]  M. Ginsberg,et al.  Dynamic regulation of integrins , 1995, Stem cells.

[42]  S. Pelech,et al.  Stimulation of Stress-activated Protein Kinase and p38 HOG1 Kinase in Murine Keratinocytes following Photodynamic Therapy with Benzoporphyrin Derivative* , 1996, The Journal of Biological Chemistry.