Objective: Cytotoxic chemotherapy of advanced breast cancer is frequently complicated by drug resistance. Our goal was to define the role of the apoptosis-regulating receptors Fas (CD95) and CD40 in the chemosensitivity of breast cancer. Methods: The sensitivity of four breast cancer cell lines to paclitaxel and mitoxantrone was evaluated using an ATP-based cell viability assay. After verification of apoptosis by annexin V staining and TUNEL assay, cell lines were characterized regarding their constitutive expression of both surface and soluble (s)Fas (CD95) and Fas ligand (Fas-L). The role of the Fas/Fas-L system and different caspases was assessed by blocking drug-mediated apoptosis with specific antibodies. Finally, the paclitaxel sensitivity of the CD40-negative cell line KS was compared to that of its CD40-positive transfectant KS-CD40. Results and Conclusion: While the cytotoxic effect of mitoxantrone did not correlate with Fas expression, the results presented here suggest some involvement of the Fas/Fas-L system in paclitaxel-induced apoptosis. Cell lines with constitutive expression of Fas/sFas demonstrated a higher sensitivity to paclitaxel than Fas-negative cells. Incubation with paclitaxel led to a measurable downregulation of the expression of both soluble and surface Fas receptor in these cells. Interestingly, stimulation of the CD40 receptor inhibited paclitaxel-induced apoptosis in the transfected cell line KS-CD40, suggesting a role of this receptor in the modulation of chemosensitivity.

1.
Barry MA, Behnke CA, Eastman A: Activation of programmed cell death (apoptosis) by cisplatin, other anticancer drugs, toxins, and hyperthermia. Biochem Pharmacol 1990;40:2352–2362.
2.
Hickman JA, Potten CS, Merritt AJ, Fisher TC: Apoptosis and cancer chemotherapy. Philos Trans R Soc Lond B Biol Sci 1994;345:319–325.
3.
Friesen C, Herr I, Krammer P, Debatin KM: Involvement of the CD95 (APO-1/Fas) receptor/ligand system in drug-induced apoptosis in leukemia cells. Nat Med 1996;5:574–577.
4.
Hamnun YA: Apoptosis and the dilemma of cancer chemotherapy. Blood 1997;89:1845–1853.
5.
Eischen CM, Kottke TJ, Martins LM, Basi GS, Tung JS, Earnshaw WC, Leibson PJ, Kaufmann SH: Comparison of apoptosis in wild-type and Fas-resistant cells: Chemotherapy-induced apoptosis is not dependent on Fas/Fas ligand interactions. Blood 1997;90:935–943.
6.
Cheng J, Zhou T, Liu C, Shapiro JP, Brauer MJ, Kiefer MC, Barr PJ, Mountz JD: Protection from Fas-mediated apoptosis by a soluble form of the Fas molecule. Science 1994;263:1759–1762.
7.
Leithauser F, Dhein J, Mechtersheimer G, Koretz K, Bruderlein S, Henne C, Schmidt A, Debatin KM, Krammer PH, Moller P: Constitutive and induced expression of apo-1, a new member of the nerve growth factor/tumor necrosis factor receptor superfamily, in normal and malignant cells. Lab Invest 1993;69:415–429.
8.
Keane MM, Ettenberg SA, Lowrey GA, Russell EK, Lipkowitz S: Fas expression and function in normal and malignant breast cell lines. Cancer Res 1996;56:4791–4798.
9.
Muschen M, Moers C, Warskulat U, Niederacher D, Betz B, Even J, Lim A, Josien R, Beckmann MW, Haussinger D: CD95 ligand expression in dedifferentiated breast cancer. J Pathol 1999;189:378–386.
10.
Mullauer L, Mosberger I, Grusch M, Rudas M, Chott A: Fas ligand is expressed in normal breast epithelial cells and is frequently upregulated in breast cancer. J Pathol 2000;190:20–30.
11.
Reimer T, Herrrnring C, Koczan D, Richter D, Gerber B, Kabelitz D, Friese K, Thiesen HJ: Fas:FasL ratio – a prognostic factor in breast carcinomas. Cancer Res 2000;60:822–828.
12.
Mottolese M, Buglioni S, Bracalenti C, Cardarelli MA, Ciabocco L, Giannarelli D, Botti C, Natali PG, Concetti A, Venanzi FM: Prognostic relevance of altered Fas (CD95)-system in human breast cancer. Int J Cancer 2000;89:127–132.
13.
Jakobson E, Jonsson G, Bjorck P, Paulie S: Stimulation of CD40 in bladder carcinoma cells inhibits anti-Fas/APO-1 (CD95)-induced apoptosis. Int J Cancer 1998;77:849–853.
14.
Sbih-Lammali F, Clausse B, Ardila-Osorio H, Guerry R, Talbot M, Havouis S, Ferradini L, Bosq J, Tursz T, Busson P: Control of apoptosis in Epstein Barr virus-positive nasopharyngeal carcinoma cells: Opposite effects of CD95 and CD40 stimulation. Cancer Res 1999;59:924–930.
15.
Wingett DG, Vestal RE, Forcier K, Hadjokas N, Nielson CP: CD40 is functionally expressed on human breast carcinomas: Variable inducibility by cytokines and enhancement of Fas-mediated apoptosis. Breast Cancer Res Treat 1998;50:27–36.
16.
Fisher DE: Apoptosis in cancer therapy: Crossing the threshold. Cell 1994;78:539–542.
17.
McCloskey DE, Armstrong DK, Jackisch C, Davidson NE: Programmed cell death in human breast cancer cells. Recent Prog Horm Res 1996;51:493–508.
18.
Muller M, Strand S, Hug H, Heinemann EM, Walczak H, Hofmann WJ, Stremmel W, Krammer PH, Galle PR: 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 1997;99:403–413.
19.
Fulda S, Los M, Friesen C, Debatin KM: Chemosensitivity of solid tumor cells in vitro is related to activation of the CD95 system. Int J Cancer 1998;76:105–114.
20.
Georgiadis MS, Russell EK, Gazdar AF, Johnson BE: Paclitaxel cytotoxicity against human lung cancer cell lines increases with prolonged exposure durations. Clin Cancer Res 1997;3:449–454.
21.
Kaufmann SH: Induction of endonucleolytic DNA cleavage in human acute myelogenous leukemia cells by etoposide, camptothecin, and other cytotoxic anticancer drugs: A cautionary note. Cancer Res 1989;49:5870–5878.
22.
Fan W: Possible mechanisms of paclitaxel-induced apoptosis. Biochem Pharmacol 1999;57:1215–1221.
23.
Hunter EM, Sutherland LA, Cree IA, Dewar JA, Preece PE, Wood RAB, Linder D, Andreotti PE: Heterogeneity of chemosensitivity in human breast carcinoma: Use of an adenosine triphosphate (ATP) chemiluminescence assay. Eur J Surg Oncol 1993;19:242–249.
24.
Young LS, Eliopoulos AG, Gallagher NJ, Dawson CW: CD40 and epithelial cells: Across the great divide. Immunol Today 1998;19:502–506.
25.
Ogretmen B, Safa AR: Down-regulation of apoptosis-related bcl-2 but not bcl-xL or bax proteins in multidrug-resistant MCF-7/Adr human breast cancer cells. Int J Cancer 1996;67:608–614.
26.
Chadderton A, Villeneuve DJ, Gluck S, Kirwan-Rhude AF, Gannon BR, Blais DE, Parissenti AM: Role of specific apoptotic pathways in the restoration of paclitaxel-induced apoptosis by valspodar in doxorubicin-resistant MCF-7 breast cancer cells. Breast Cancer Res Treat 2000;59:231–244.
27.
Biswas RS, Cha HJ, Hardwick JM, Srivastava RK: Inhibition of drug-induced Fas ligand transcription and apoptosis by Bcl-XL. Mol Cell Biochem 2001;225:7–20.
28.
Ferreira CG, Tolis C, Span SW, Peters GJ, van Lopik T, Kummer AJ, Pinedo HM, Giaccone G: Drug-induced apoptosis in lung cancer cells is not mediated by the Fas/FasL (CD95/APO1) signalling pathway. Clin Cancer Res 2000;6:203–212.
29.
Odoux C, Albers A, Amoscato AA, Lotze MT, Wong MKK: TRAIL, FasL and a blocking anti-DR5 antibody augment paclitaxel-induced apoptosis in human non-small-cell lung cancer. Int J Cancer 2002;97:458–465.
30.
Friesen C, Fulda S, Debatin KM: Deficient activation of the CD95 (APO-1/Fas) receptor system in drug-resistant cells. Leukemia 1997;11:1833–1841.
31.
Fulda S, Sieverts H, Friesen C, Herr I, Debatin KM: The CD95 (APO-1/Fas) system mediates drug-induced apoptosis in neuroblastoma cells. Cancer Res 1997;57:3823–3829.
32.
McGahon AJ, Costa PA, Daly L, Cotter TG: Chemotherapeutic drug-induced apoptosis in human leukemia cells is independent of the Fas (APO-1/CD95) receptor/ligand system. Br J Haematol 1998;101:539–547.
33.
Chlichlia K, Peter ME, Scaffidi C, Bucur M, Krammer PH, Schirrmacher V, Umansky V: Caspase activation is required for nitric oxide-mediated, CD95 (APO-1/Fas)-dependent and independent apoptosis in human neoplastic lymphoid cells. Blood 1998;91:4311–4320.
34.
Batrla R, Linnebacher M, Rudy W, Stumm S, Wallwiener D, Gückel B: CD40-expressing carcinoma cells induce down-regulation of CD40-ligand (CD154) and impair T cell function. Cancer Res 2002;62:2052–2057.
35.
Mizutani Y, Yoshida O, Bonavida B: Prognostic significance of soluble Fas in the serum of patients with bladder cancer. J Urol 1998;160:571–576.
36.
Strand S, Hoffman WJ, Hug H, Müller M, Otto G, Strand D, Mariani SM, Stremmel W, Krammer PH, Galle PR: Lymphocyte apoptosis induced by CD95 (APO-1/Fas) ligand-expressing tumor cells – a mechanism of immune evasion? Nat Med 1996;2:1361–1366.
37.
van Kooten C, Banchereau J: Functions of CD40 on B cells, dendritic cells and other cells. Curr Opin Immunol 1997;9:330–337.
38.
Suda T, Nagata S: Purification and characterization of the Fas-ligand that induces apoptosis. J Exp Med 1994;179:873–879.
39.
Keane MM, Ettenberg SA, Nau MM, Russell EK, Lipkowitz S: Chemotherapy augments TRAIL-induced apoptosis in breast cell lines. Cancer Res 1999;59:734–741.
40.
Guttierez LS, Eliza M, Niven-Fairchild T, Naftolin F, Mor G: The Fas/Fas-ligand system: A mechanism for immune evasion in human breast carcinomas. Breast Cancer Res Treat 1999;54:245–253.
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