title: TIMP-1-GPI in combination with hyperthermic treatment of melanoma increases sensitivity to FAS-mediated apoptosis.
abstract: Resistance to apoptosis is a prominent feature of malignant melanoma. Hyperthermic therapy can be an effective adjuvant treatment for some tumors including melanoma. We developed a fusion protein based on the tissue inhibitor of matrix metalloproteinase-1 linked to a glycosylphosphatidylinositol anchor (TIMP-1-GPI). The TIMP-1-GPI-fusion protein shows unique properties. Exogenous administration of TIMP-1-GPI can result in transient morphological changes to treated cells including modulation of proliferation and decreased resistance to apoptosis. The effect of TIMP-1-GPI on the biology of melanoma in the context of a defined hyperthermic dose was evaluated in vitro. Clonogenic assays were used to measure cell survival. Gelatinase zymography determined secretion of MMP-2 and MMP-9. Monoclonal antibody against FAS/CD95 was applied to induce apoptosis. The expression of pro- and anti-apoptotic proteins and the secretion of immunoregulatory cytokines were then evaluated using Western blot and ELISA. TIMP-1-GPI combined with a sub-lethal hyperthermic treatment (41.8 degrees C for 2 h) suppressed tumor cell growth capacity as measured by clonogenic assay. The co-treatment also significantly suppressed tumor cell proliferation, enhanced FAS receptor surface expression increased tumor cell susceptibility to FAS-mediated killing. The increased sensitivity to FAS-induced apoptosis was linked to alterations in the apoptotic mediators Bcl-2, Bax, Bcl-XL and Apaf-1. The agent works in concert with sub-lethal hyperthermic treatment to render melanoma cells sensitive to FAS killing. The targeted delivery of TIMP-1-GPI to tumor environments in the context of regional hyperthermic therapy could be optimized through the use of thermosensitive liposomes.
authors.affiliation: Medizinische Poliklinik, Ludwig-Maximilians-Universität München, Munich, Germany.
authors.forename: Roghieh
authors.initials: R
authors.lastname: Djafarzadeh
authors.forename: Valeria
authors.initials: V
authors.lastname: Milani
authors.forename: Nicole
authors.initials: N
authors.lastname: Rieth
authors.forename: Irene
authors.initials: I
authors.lastname: von Luettichau
authors.forename: Petra S
authors.initials: PS
authors.lastname: Skrablin
authors.forename: Monika
authors.initials: M
authors.lastname: Hofstetter
authors.forename: Elfriede
authors.initials: E
authors.lastname: Noessner
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Nelson
mesh_terms: D017209:Apoptosis; D049109:Cell Proliferation; D002469:Cell Separation; D002470:Cell Survival; D005434:Flow Cytometry; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D006801:Humans; D006979:Hyperthermia, Induced; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D008545:Melanoma / Q000628:therapy*; D012878:Skin Neoplasms / Q000473:pathology* / Q000628:therapy*; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000378:metabolism*; D016896:Treatment Outcome; D019014:fas Receptor / Q000378:metabolism*
chemical_list: D017261:Glycosylphosphatidylinositols; D019715:Tissue Inhibitor of Metalloproteinase-1; D019014:fas Receptor; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9