Abstract
The pronounced resistance of human renal cell carcinoma (RCC) to anticancer-induced apoptosis has primarily been related to the expression of P-glycoprotein and effective drug detoxification mechanisms. Because the CD95 system has recently been identified as a key mediator of anticancer drug-induced apoptosis, we analysed the contribution of the CD95 system to chemotherapy-induced apoptosis in four newly established RCC cell lines. Here, we demonstrate that all RCC cell lines expressed CD95-receptor and -ligand. Exposure to agonistic anti-CD95 antibodies resulted in induction of apoptosis and significant (P< 0.05) reduction of cell number in three out of four cell lines, indicating that the essential components for CD95-mediated apoptosis were present and functionally intact in the majority of these RCC cell lines. Moreover, treatment of cultures with bleomycin or topotecan, a novel topoisomerase I inhibitor with little substrate affinity for P-glycoprotein, led to induction of apoptosis and significant (P< 0.05) dose-dependent reduction of cell number in all RCC cell lines. Both anticancer drugs also induced upregulation of CD95 ligand expression in all cell lines. Additionally, augmentation of CD95 receptor expression was found in three RCC cell lines, including one p53-mutated cell line, whereas another p53-mutated cell line showed no or only a weak CD95 receptor upregulation after exposure to topotecan or bleomycin, respectively. Despite this upregulation of CD95 receptor and ligand, antagonistic antibodies directed against CD95 receptors or ligands could not inhibit induction of apoptosis by topotecan and bleomycin in any cell line. Thus, although a functionally intact CD95 signalling cascade is present in most RCC cell lines, the anticancer drugs topotecan and bleomycin that induce upregulation of CD95 receptor and ligand fail to effectively activate CD95-mediated apoptosis. This deficient activation of CD95-mediated apoptosis might be an important additional factor for the multidrug resistance phenotype of human RCCs. © 2000 Cancer Research Campaign
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References
Baldini N (1997) Multidrug resistance – a multiplex phenomenon. Nat Med 3: 78–380
Caignard A, Guillard M, Cai C, Asselin-Paturel C, Carayol G and Chouaib S (1996) The renal cell carcinoma lysis by a specific cytotoxic T cell clone is independent of the Fas/Fas-L cytotoxic pathway. Tissue Antigens 48: 295–300
Cascino I, Fiucci G, Pappoff G and Ruberti G (1995) Three functional soluble forms of the human apoptosis-inducing Fas molecule are produced by alternative splicing. J Immunol 154: 2700–2713
El-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW and Vogelstein B (1993) WAF-1, a potential mediator of p53 tumor suppression. Cell 75: 817–825
Friesen C, Herr I, Krammer PH and Debatin KM (1996) Involvement of the CD95 (APO-1/Fas) receptor/ligand system in drug-induced apoptosis in leukemia cells. Nat Med 2: 574–577
Friesen C, Fulda S and Debatin KM (1997) Deficient activation of the CD95 (APO-1/Fas) system in drug-resistant cells. Leukemia 11: 1833–1841
Fulda S, Sieverts H, Friesen C, Herr I and Debatin KM (1997) The CD95 (APO-1/Fas) system mediates drug-induced apoptosis in neuroblastoma cells. Cancer Res 57: 3823–3829
Fulda S, Los M, Friesen C and Debatin KM (1998) Chemosensitivity of solid tumor cells in vitro is related to activation of the CD95 system. Int J Cancer 76: 105–114
Gerharz CD, Moll R, Störkel S, Ramp U, Thoenes W and Gabbert HE (1993) Ultrastructural appearance and cytoskeletal architecture of the clear cell, chromophilic and chromophobe cell variants of human renal cell carcinoma in vivo and in vitro. Am J Pathol 142: 851–859
Gerharz CD, Ramp U, Olert J, Moll R, Störkel S, Marx N and Gabbert HE (1994) Cytomorphological, cytogenetic, and molecular biological characterization of four new human renal carcinoma cell lines of the clear cell type. Virchows Arch B 424: 403–409
Gerharz CD, Moll R, Störkel S, Ramp U, Hildebrandt B, Molsberger G, Koldovsky P and Gabbert HE (1995) Establishment and characterization of two divergent cell lines derived from a human chromophobe renal cell carcinoma. Am J Pathol 146: 953–962
Gerharz CD, Hilderbrandt B, Moll R, Ramp U, Sarbia M, Störkel S, Koldovsky P and Gabbert HE (1996) Chromophilic renal cell carcinoma: cytomorphological and cytogenetic characterization of four permanent cell lines. Br J Cancer 74: 1605–1614
Harris CC and Hollstein M (1993) Clinical implications of the p53 tumor-suppressor gene. New Engl J Med 329: 1318–1327
Hendricks CB, Rowinsky EK, Grochow LB, Donehower RC and Kaufmann SH (1992) Effect of P-glycoprotein expression on the accumulation and cytotoxicity of topotecan (SK&F 104864), a new camptothecin analogue. Cancer Res 52: 2268–2278
Herben VMM, ten Bokkel Huinink WW and Beijnen JH (1996) Clinical pharmacokinetics of topotecan. Clin Pharmacokinet 31: 85–102
Hetts SW (1998) To die or not to die. An overview of apoptosis and its role in disease. JAMA 279: 300–307
Hickman JA (1992) Apotosis induced by anti-cancer agents. Cancer Metastasis Rev 11: 121–139
Homma Y and Aso Y (1994) Effect of α-Interferon alone and combined with other antineoplastic agents on renal cell carcinoma determined by the tetrazolium microculture assay. Eur Urol 25: 164–170
Horie S, Kano M, Higashihara E, Moriyama N, Tanaka E, Hirose A, Kakizoe T and Kawabe K (1997) Expression of Fas in renal cell carcinoma. Jpn J Clin Oncol 27: 384–388
Keane MM, Ettenberg SA, Lowrey GA, Russel EK and Lipkowitz S (1996) Fas expression and function in normal and malignant breast cell lines. Cancer Res 56: 4791–4798
Kischkel FC, Hellbardt S, Behrmann I, Germer M, Pawlita M, Krammer PH and Peter ME (1995) Cytotoxicity-dependent APO-1 (Fas/CD95)-associated proteins form a death-inducing signalling complex (DISC) with the receptor. EMBO J 14: 5579–5588
Krammer PH (1997) The tumor strikes back. New data on expression of the CD95 (APO-1/Fas) receptor/ligand system may cause paradigm changes in our view on drug treatment and tumor immunology. Cell Death Differentiation 4: 362–364
Mickisch GH, Bier H, Bergler W, Bak M, Tschada R and Alken P (1990) P-170 glycoprotein, glutathione and associated enzymes in relation to chemoresistance of primary human renal cell carcinomas. Urol Int 45: 170–176
Mitsui I, Kumazawa E, Hirota Y, Aonuma M, Sugimori M, Ohsuki S, Uoto K, Ejima A, Terasawa H and Sato K (1995) A new water-soluble camptothecin derivate, DX-8951f, exhibits potent antitumor activity against human tumors in vitro and in vivo. Jpn J Cancer Res 86: 776–782
Miyake H, Hara I, Gohji K, Arakawa S and Kamidono S (1998) p53 modulation of Fas/APO-1 mediated apoptosis in a human renal cell carcinoma cell line. Int J Oncol 12: 469–473
Moll UM and Schramm LM (1998) p53 – an acrobat in tumorigenesis. Crit Rev Oral Biol Med 9: 23–37
Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65: 55–64
Mulders P, Figlin R, deKernion JB, Wiltrout R, Linehan M, Parkinson D, deWolf W and Belldegrun A (1997) Renal cell carcinoma: recent progress and future directions. Cancer Res 57: 5189–5195
Müller M, Strand S, Hug H, Heinemann EM, Walczak H, Hofmann WJ, Stremmel W, Krammer PH and Galle PR (1997) Drug-induced apoptosis in hepatoma cells is mediated by the CD95 (APO-1/Fas) receptor/ligand system and involves activation of wild-type p53. J Clin Invest 99: 403–413
Müller M, Wilder S, Bannasch D, Israeli D, Lehlbach K, Li-Weber M, Friedman SL, Galle PR, Stremmel W, Oren M and Krammer PH (1998) p53 activates the CD95 (APO-1/Fas) gene in response to DNA damage by anticancer drugs. J Exp Med 11: 2033–2045
Nagata S (1994) Apoptosis regulated by a death factor and its receptor: Fas ligand and Fas. Philos Trans R Soc Lond B Biol Sci 345: 281–287
Nakajima H, Yamada N and Takiguchi M (1998) Fas-independent apoptosis of T cells via killer cell inhibitory receptors. Int Immunol 10: 85–90
Natoli G, Ianni A, Constanzo A, De Petrillo G, Ilari I, Chirillo P, Balsano C and Levrero M (1995) Resistance to Fas-mediated apoptosis in human hepatoma cells. Oncogene 11: 1157–1164
Nonomura N, Miki T, Yokoyama M, Imazu T, Takada T, Takeuchi S, Kanno N, Nihimura K, Kojima Y and Okuyama A (1996) Fas/APO-1-mediated apoptosis of human renal cell carcinoma. Biochem Biophys Res Commun 229: 945–951
O’Connell J, O’Sullivan GC, Collins JK and Shanahan F (1996) The Fas counterattack: Fas-mediated T cell killing by colon cancer cells expressing Fas ligand. J Exp Med 184: 1075–1082
Owen-Schaub LB, Zhang W, Cusack JC, Angelo LS, Santee SM, Fujiwara T, Roth JA, Deisseroth AB, Zhang WW, Kruzel E and Radinsky R (1995) Wild-type human p53 and a temperature-sensitive mutant induce FAS/APO-1 expression. Mol Cell Biol 15: 3032–3040
Peter ME, Dhen I, Ehret A, Helbardt S, Walczak H, Moldenhauer G and Krammer PH (1995) APO-1 (CD95)-dependent and independent antigen-receptor-induced apoptosis in human T and B cell lines. Int Immunol 7: 1873–1877
Ramp U, Jaquet K, Reinecke P, Nitsch T, Gabbert HE and Gerharz CD (1997) Acquisition of TGF-β1resistance: an important progression factor in human renal cell carcinoma. Lab Invest 76: 739–749
Reiter RE, Anglard P, Liu S, Gnarra JR and Linnehan WM (1993) Chromosome 17p deletions and p53 mutations in renal cell carcinoma. Cancer Res 53: 3092–3097
Scaffidi C, Fulda S, Srinivasan A, Friesen C, Li F, Tomaselli KJ, Debatin KM, Krammer PH and Peter ME (1998) Two CD95 (APO-1/Fas) signalling pathways. EMBO J 17: 1675–1687
Scheithauer W, Clark GM, Salmon SE, Dorda W, Shoemaker RH and von Hoff DD (1986) Model for estimation of clinically achievable plasma concentrations for investigational anticancer drugs in man. Cancer Treatment Reports 70: 1379–1382
Schulze-Osthoff K, Ferrari D, Los M, Wesselborg S and Peter ME (1998) Apoptosis signalling by death receptors. Eur J Biochem 254: 439–459
Sorensen M, Sehested M and Jensen PB (1995) Characterisation of a human small-cell lung cancer cell line resistant to the DNA topoisomerase I-directed drug topotecan. Br J Cancer 72: 399–404
Strand S, Hofmann WJ, Hug H, Müller M, Otto G, Strand D, Mariani SM, Stremmel W, Krammer PH and Galle PR (1996) Lymphocyte apoptosis induced by CD95 (APO-1/Fas) ligand-expressing tumor cells – a mechanism of immune evasion?. Nat Med 2: 1361–1366
Tomita Y, Kawasaki T, Bilim V, Takeda M and Takahashi K (1996) Tetrapeptide DEVD-aldehyde or YVAD-chloromethylketone inhibits Fas/APO-1 (CD95)-mediated apoptosis in renal cancer cells. Int J Cancer 68: 132–135
Waldman T, Kinzler KW and Vogelstein B (1995) p21 is necessary for the p53-mediated G1 arrest in human cancer cells. Cancer Res 55: 5187–5190
Webb JL (1963) Effects of more than one inhibitor. Enzyme and Metabolic Inhibitors 1: 487–512, Academic Press: New York
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Ramp, U., Dejosez, M., Mahotka, C. et al. Deficient activation of CD95 (APO-1/ Fas)-mediated apoptosis: a potential factor of multidrug resistance in human renal cell carcinoma. Br J Cancer 82, 1851–1859 (2000). https://doi.org/10.1054/bjoc.2000.1155
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DOI: https://doi.org/10.1054/bjoc.2000.1155
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