Abstract
Glioblastoma multiforme (GBM) is the most frequent malignant brain tumor in adults and is invariably fatal. We have investigated the effect of cyclo-(Arg-Gly-Asp-D-Phe-Val) (cRGDfV) peptide on survival of human malignant glioma cells in vitro and in vivo. Immunofluorescent analyses revealed the presence of αVβ3 integrin on U-87MG and U-373MG cells, but minimal expression on U-251MG cells. Treatment of U-87MG and U-373MG cells in vitro with cRGDfV (20 µg/ml), but not the linear peptide, resulted in the appearance of rounded and loosely attached cells with subsequent cell death. By comparison, neither this cyclic peptide nor its linear homolog had any significant effect on growth and morphology of U-251MG cells. The death of cRGDfV-treated (20 µg/ml) glioma cells was blocked by pretreatment (10 µM) of cells with DEVD-FMK and LEHD-FMK, inhibitors of caspase-3 and caspase-9, respectively. Moreover, when glioma cells grown as spheroids were treated with cRGDfV (50 µg/ml), spheroid formation was markedly reduced. Further, treatment of intracranial U-87MG tumors in scid mice with cyclic peptide significantly (p<0.001) prolonged their survival. These results indicated (i) that cRGDfV induced apoptosis of human glioma cells by binding αVβ3 integrin expressed on their cell surfaces and (ii) that cRGDfV may be an effective and non-toxic direct anti-tumor therapy for αVβ3-expressing GBMs.
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References
Cavenee WK, Bigner DD, Newcomb EW, Paulus W, Kleihues P: Diffuse astrocytomas. In: Kleihues P, Cavenee WK (eds). Pathology and Genetics of Tumours of the Nervous System. IARC Press, Lyon, France, 1997, pp 2-9
Mahaley MS, Jr, Mettlin C, Natarajan N, Laws ER, Peace BB: National survey of patterns of care for brain-tumor patients. J Neurosurg 71: 826-836, 1989
Walker MD, Green SB, Byar DP, Alexander E, Jr, Balzdorf U, Brooks WH, Hunt WE, MacCarty CS, Mahaley MS, Jr, Mealey J, Jr, Owens G, Ransohoff JI, Robertson JT, Shapiro WR, Smith KR, Jr, Wilson CB, Strike TA: Randomized comparisons of radiotherapy and nitrosoureas for the treatment of malignant glioma after surgery. N Engl J Med 303: 1323-1329, 1980
Brooks PC, Montgomery AMP, Rosenfeld M, Reisfeld RA, Hu T, Klier G, Cheresh DA: Integrin αvβ3 antagonists promote tumor regression by inducing apoptosis of angiogenic blood vessels. Cell 79: 1157-1164, 1994
Brooks PC, Clark RAF, Cheresh DA: Requirement of vascular integrin αvβ3for angiogenesis. Science 264: 569-571, 1994
Ruegg C, Yilmaz A, Bieler G, Bamat J, Chaubert P, Lejeune FJ: Evidence for the involvement of endothelial cell integrin αvβ3 in the disruption of the tumor vasculature induced by TNF and IFN-γ. Nat Med 4: 408-414, 1998
Friedlander M, Brooks PC, Shaffer RW, Kincaid CM, Varner JA, Cheresh DA: Definition of two angiogenic pathways by distinct αvintegrins. Science 270: 1500-1502, 1995
Chothia C, Jones EY: The molecular structure of cell adhesion molecules. Annu Rev Biochem 66: 823-862, 1997
Ruoslahti E: RGD and other recognition sequences for integrins. Annu Rev Cell Dev Biol 12: 697-715, 1996
Felding-Habermann B, Cheresh DA: Vitronectin and its receptors. Curr Opin Cell Biol 5: 864-868, 1993
Chatterjee S, Bradac J, Hunter E: Effect of monensin on Mason-Pfizer monkey virus glycoprotein synthesis. J Virol 44: 1003-1012, 1982
Chambers R, Gillespie GY, Soroceanu L, Andreansky S, Chatterjee S, Chou J, Roizman B, Whitley RJ: Comparison of genetically engineered herpes simplex viruses for the treatment of brain tumors in a scid mouse model of human malignant glioma. Proc Natl Acad Sci USA, 92: 1411-1415, 1995
Salvesen GS, Dixit VM: Caspases: intracellular signaling by proteolysis. Cell 91: 443-446, 1997
Cohen GM: Caspases: the executioners of apoptosis. Biochem J 326: 1-16, 1997
Janicke RU, Sprengart ML, Wati MR, Porter AG: Caspase-3 is required for DNA fragmentation and morphological changes associated with apoptosis. J Biol Chem 273: 9357-9360, 1998
Cardone MH, Roy N, Stennicke HR, Salvesen GS, Franke TF, Stanbridge E, Frisch S, Reed JC: Regulation of cell death protease caspase-9 by phosphorylation. Science 282: 1318-1321, 1998
Sakahira H, Enari M, Nagata S: Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis. Nature 391: 96-99, 1998
Gladson CL, Cheresh DA: Glioblastoma expression of vitronectin and the αvβ3 integrin: adhesion mechanism for transformed glial cells. J Clin Invest 88: 1924-1932, 1991
Gladson C, Wilcox JN, Sanders L, Gillespie GY, Cheresh DA: Cerebral microenvironment influences expression of the vitronectin gene in astrocytic tumors. J Cell Science 108: 947-956, 1995
Wikstrand CJ, Bigner SH, Bigner DD: Demonstration of complex antigenic heterogenecity in a human glioma cell line and eight derived clones by specific monoclonal antibodies. Cancer Res 43: 3327-3334, 1983
Kaufmann SH, Desnoyers S, Ottaviano Y, Davidson NE, Poirier GG: Specific proteolytic cleavage of poly (ADP-Ribose) polymerase: an early marker of chemotherapyinduced apoptosis. Cancer Res 53: 3976-3985, 1993
Satoh MS, Lindahl T: Role of poly (ADP-Ribose) formation in DNA repair. Nature 356: 356-358, 1992
Zou H, Henzel WJ, Liu X, Lutschg A, Wang X: Apaf-1, a human protein homologous to C. elegans CED-4, participates in cytochrome c-dependent activation of caspase-3. Cell 90: 405-413, 1997
Stromblad S, Becker J, Yerba M, Brooks P, Cheresh D: Suppression of p53 activity and p21 expression by vascular cell integrin αvβ3 during angiogenesis. J Clin Invest 98: 426-433, 1996
Gomez-Manzano C, Fueyo J, Kyritis AP, McDonnell TJ, Steck PA, Levin VA, Yung W: Characterization of p53 and p21 functional interactions in glioma cells en route to apoptosis. J Natl Cancer Inst 89: 1036-1044, 1997
Vuori K, Ruoslahti E: Association of insulin receptor substrate-1 with integrins. Science 266: 1576-1578, 1994
Schneller M, Vuori K, Ruoslahti E: αvβ3 integrin associates with activated insulin and PDGF-? receptors and potentiates the biological activity of PDGF. EMBO J 16: 5600-5607, 1997
Nister M, Clasesson-Welch L, Enksson A, Heldin CH, Westermark B: Differential expression of platelet-derived growth factor receptors in human malignant glioma cell lines. J Biol Chem 266: 16755-16763, 1991
Glick RP, Unterman TG, Hollis R: Radioimmunoassay of insulin-like growth factors in cyst fluid of central nervous system tumors. J Neurosurg 74: 972-978, 1991
Rubin R, Baserga R: Insulin-like growth factor-I receptor: its role in cell proliferation, apoptosis and tumorigenicity. Lab Invest 73: 311-331, 1995
Yang B-C, Chang H-M, Wang Y-S, Chen R-F, Lin S-J: Transient induction of apoptosis in serum-starved glioma cells by insulin and IGF-I. Biochim Biophys Acta 1314: 83-92, 1996
Burgaud JL, Resnicoff M, Baserga R: Mutant IGF-I receptors as dominant negatives for growth and transformation. Biochem Biophys Res Commun 214: 475-481, 1995
Resnicoff M, Li W, Basak S, Herlyn D, Baserga R, Rubin R: Inhibition of rat C6 glioblastoma tumor growth by expression of insulin-like growth factor I receptor antisense mRNA. Cancer Immunol Immunother 42: 64-68, 1996
Kulik G, Kippel A, Weber MJ: Antiapoptotic signaling by the insulin-like growth factor-I receptor, phosphatidylinositol 3-kinase and Akt. Mol Cell Biol 17: 1595-1606, 1997
Saitoh Y, Kurastsu J, Takeshima H, Yamamoto S, Ushio Y: Expression of osteopontin in human gliomas: its correlation with malignancy. Lab Invest 72: 55-63, 1995
Tucker MA, Chang P, Prince CW, Gillespie GY, Mapstone TB: TPA-mediated regulation of osteopontin in human malignant glioma cells. Anticancer Res 18: 807-812, 1998
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Chatterjee, S., Matsumura, A., Schradermeier, J. et al. Human Malignant Glioma Therapy Using Anti-αVβ3 Integrin Agents. J Neurooncol 46, 135–144 (2000). https://doi.org/10.1023/A:1006444300504
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DOI: https://doi.org/10.1023/A:1006444300504