Abstract
Tumor necrosis factor-related apoptosis inducing ligand (TRAIL) shows promise as a chemotherapeutic agent. However, many human cancer cells are resistant to killing by TRAIL. We have previously demonstrated that reovirus infection increases the susceptibility of human lung (H157) and breast (ZR75-1) cancer cell lines to TRAIL-induced apoptosis. We now show that reovirus also increases the susceptibility of human ovarian cancer cell lines (OVCAR3, PA-1 and SKOV-3) to TRAIL-induced apoptosis. Reovirus-induced increases in susceptibility of OVCAR3 cells to TRAIL require virus uncoating and involve increased activation of caspases 3 and 8. Reovirus infection results in the down-regulation of cFLIP (cellular FLICE inhibitory protein) in OVCAR3 cells. Down-regulation of cFLIP following treatment of OVCAR3 cells with antisense cFLIP oligonucleotides or PI3 kinase inhibition also increases the susceptibility of OVCAR3 cells to TRAIL-induced apoptosis. Finally, over-expression of cFLIP blocks reovirus-induced sensitization of OVCAR3 cells to TRAIL-induced apoptosis. The combination of reovirus and TRAIL thus represents a promising new therapeutic approach for the treatment of ovarian cancer.
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Landis SH, Murray T, Bolden S, Wingo PA (1999) Cancer statistics, 1999. CA Cancer J Clin 49:8–31, 1
Partridge EE, Barnes MN (1999) Epithelial ovarian cancer: Prevention, diagnosis, and treatment. CA Cancer J Clin 49:297–20
Ashkenazi A, Dixit VM (1998) Death receptors: Signaling and modulation. Science 281:1305–308
Hao C, Beguinot F, Condorelli G, et al (2001) Induction and intracellular regulation of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) mediated apotosis in human malignant glioma cells. Cancer Res 61:1162–170
Chang DW, Xing Z, Pan Y, et al (2002) c-FLIP(L) is a dual function regulator for caspase-8 activation and CD95-mediated apoptosis. EMBO J 21:3704–714
Ashkenazi A, Pai RC, Fong S, et al (1999) Safety and antitumor activity of recombinant soluble Apo2 ligand. J Clin Invest 104:155–62.
Walczak H, Miller RE, Ariail K, et al (1999) Tumoricidal activity of tumor necrosis factor-related apoptosis-inducing ligand in vivo. Nat Med 5:157–63
Wiley SR, Schooley K, Smolak PJ, et al (1995) Identification and characterization of a new member of the TNF family that induces apoptosis. Immunity 3:673–82
Pitti RM, Marsters SA, Ruppert S, Donahue CJ, Moore A, Ashkenazi A (1996) Induction of apoptosis by Apo-2 ligand, a new member of the tumor necrosis factor cytokine family. J Biol Chem 271:12687–2690
Bodmer JL, Holler N, Reynard S, et al (2000) TRAIL receptor-2 signals apoptosis through FADD and caspase-8. Nat Cell Biol 2:241–43
Kischkel FC, Hellbardt S, Behrmann I, et al (1995) Cytotoxicity-dependent APO-1 (Fas/CD95)-associated proteins form a death-inducing signaling complex (DISC) with the receptor. EMBO J 14:5579–588
Muzio M, Stockwell BR, Stennicke HR, Salvesen GS, Dixit VM (1998) An induced proximity model for caspase-8 activation. J Biol Chem 273:2926–930
Boatright KM, Renatus M, Scott FL, et al (2003) A unified model for apical caspase activation. Mol Cell 11:529–41
Degli-Esposti MA, Smolak PJ, Walczak H, et al (1997) Cloning and characterization of TRAIL-R3, a novel member of the emerging TRAIL receptor family. J Exp Med 186:1165–170
Degli-Esposti MA, Dougall WC, Smolak PJ, Waugh JY, Smith CA, Goodwin RG (1997) The novel receptor TRAIL-R4 induces NF-kappaB and protects against TRAIL-mediated apoptosis, yet retains an incomplete death domain. Immunity 7:813–20
Marsters SA, Sheridan JP, Pitti RM, et al (1997) A novel receptor for Apo2L/TRAIL contains a truncated death domain. Curr Biol 7:1003–006
Sheridan JP, Marsters SA, Pitti RM, et al (1997) Control of TRAIL-induced apoptosis by a family of signaling and decoy receptors. Science 277:818–21
Emery JG, McDonnell P, Burke MB, et al (1998) Osteoprotegerin is a receptor for the cytotoxic ligand TRAIL. J Biol Chem 273:14363–4367
Holen I, Croucher PI, Hamdy FC, Eaton CL (2002) Osteoprotegerin (OPG) is a survival factor for human prostate cancer cells. Cancer Res 62:1619–623
Neville-Webbe HL, Cross NA, Eaton CL, et al (2004) Osteoprotegerin (OPG) produced by bone marrow stromal cells protects breast cancer cells from TRAIL-induced apoptosis. Breast Cancer Res Treat 86:269–79
Shipman CM, Croucher PI (2003) Osteoprotegerin is a soluble decoy receptor for tumor necrosis factor-related apoptosis-inducing ligand/Apo2 ligand and can function as a paracrine survival factor for human myeloma cells. Cancer Res 63:912–16
Bonavida B, Ng CP, Jazirehi A, Schiller G, Mizutani Y (1999) Selectivity of TRAIL-mediated apoptosis of cancer cells and synergy with drugs: The trail to non-toxic cancer therapeutics (review). Int J Oncol 15:793–02
Eggert A, Grotzer MA, Zuzak TJ, Wiewrodt BR, Ikegaki N, Brodeur GM (2000) Resistance to TRAIL-induced apoptosis in neuroblastoma cells correlates with a loss of caspase-8 expression. Med Pediatr Oncol 35:603–07
Grotzer MA, Eggert A, Zuzak TJ, et al (2000) Resistance to TRAIL-induced apoptosis in primitive neuroectodermal brain tumor cells correlates with a loss of caspase-8 expression. Oncogene 19:4604–610
Hu WH, Johnson H, Shu HB (2000) Activation of NF-kappaB by FADD, Casper, and caspase-8. J Biol Chem 275:10838–0844
Deng Y, Lin Y, Wu X (2002) TRAIL-induced apoptosis requires Bax-dependent mitochondrial release of Smac/DIABLO. Genes Dev 16:33–5
Fulda S, Meyer E, Debatin KM (2002) Inhibition of TRAIL-induced apoptosis by Bcl-2 overexpression. Oncogene 21:2283–294
Hinz S, Trauzold A, Boenicke L, et al (2000) Bcl-XL protects pancreatic adenocarcinoma cells against CD95- and TRAIL-receptor-mediated apoptosis. Oncogene 19:5477–486
Kim K, Fisher MJ, Xu SQ, el Deiry WS (2000) Molecular determinants of response to TRAIL in killing of normal and cancer cells. Clin Cancer Res 6:335–46
Siegmund D, Hadwiger P, Pfizenmaier K, Vornlocher HP, Wajant H (2002) Selective inhibition of FLICE-like inhibitory protein expression with small interfering RNA oligonucleotides is sufficient to sensitize tumor cells for TRAIL-induced apoptosis. Mol Med 8:725–32
Cuello M, Ettenberg SA, Nau MM, Lipkowitz S (2001) Synergistic induction of apoptosis by the combination of trail and chemotherapy in chemoresistant ovarian cancer cells. Gynecol Oncol 81:380–90
Siervo-Sassi RR, Marrangoni AM, Feng X, et al (2003) Physiological and molecular effects of Apo2L/TRAIL and cisplatin in ovarian carcinoma cell lines. Cancer Lett 190:61–2
Lane D, Cartier A, L’Esperance S, Cote M, Rancourt C, Piche A (2004) Differential induction of apoptosis by tumor necrosis factor-related apoptosis-inducing ligand in human ovarian carcinoma cells. Gynecol Oncol 93:594–04
Vignati S, Codegoni A, Polato F, Broggini M (2002) Trail activity in human ovarian cancer cells: Potentiation of the action of cytotoxic drugs. Eur J Cancer 38:177–83
Tomek S, Horak P, Pribill I, et al (2004) Resistance to TRAIL-induced apoptosis in ovarian cancer cell lines is overcome by co-treatment with cytotoxic drugs. Gynecol Oncol 94:107–14
Horak P, Pils D, Kaider A, et al (2005) Perturbation of the tumor necrosis factor–related apoptosis-inducing ligand cascade in ovarian cancer: Overexpression of FLIPL and deregulation of the functional receptors DR4 and DR5. Clin Cancer Res 11:8585–591
Krueger A, Schmitz I, Baumann S, Krammer PH, Kirchhoff S (2001) Cellular FLICE-inhibitory protein splice variants inhibit different steps of caspase-8 activation at the CD95 death-inducing signaling complex. J Biol Chem 276:20633–0640
Chang DW, Xing Z, Pan Y, et al (2002) c-FLIP(L) is a dual function regulator for caspase-8 activation and CD95-mediated apoptosis. EMBO J 21:3704–714
Micheau O, Thome M, Schneider P, et al (2002) The long form of FLIP is an activator of caspase-8 at the Fas death-inducing signaling complex. J Biol Chem 277:45162–5171
Boatright KM, Deis C, Denault JB, Sutherlin DP, Salvesen GS (2004) Activation of caspases-8 and -10 by FLIP(L). Biochem J 382:651–57
Strong JE, Lee PW (1996) The v-erbB oncogene confers enhanced cellular susceptibility to reovirus infection. J Virol 70:612–16
Strong JE, Coffey MC, Tang D, Sabinin P, Lee PW (1998) The molecular basis of viral oncolysis: Usurpation of the Ras signaling pathway by reovirus. EMBO J 17:3351–362
Coffey MC, Strong JE, Forsyth PA, Lee PW (1998) Reovirus therapy of tumors with activated Ras pathway. Science 282:1332–334
Hirasawa K, Nishikawa SG, Norman KL, Alain T, Kossakowska A, Lee PW (2002) Oncolytic reovirus against ovarian and colon cancer. Cancer Res 62:1696–701
Alain T, Hirasawa K, Pon KJ, et al (2002) Reovirus therapy of lymphoid malignancies. Blood 100:4146–153
Norman KL, Coffey MC, Hirasawa K, et al (2002) Reovirus oncolysis of human breast cancer. Hum Gene Ther 13:641–52
Wilcox ME, Yang W, Senger D, et al (2001) Reovirus as an oncolytic agent against experimental human malignant gliomas. J Natl Cancer Inst 93:903–12
Ikeda Y, Nishimura G, Yanoma S, Kubota A, Furukawa M, Tsukuda M (2004) Reovirus oncolysis in human head and neck squamous carcinoma cells. Auris Nasus Larynx 31:407–12
Tyler KL, Squier MK, Brown AL, et al (1996) Linkage between reovirus-induced apoptosis and inhibition of cellular DNA synthesis: Role of the S1 and M2 genes. J Virol 70:7984–991
Connolly JL, Dermody TS (2002) Virion disassembly is required for apoptosis induced by reovirus. J Virol 76:1632–641
Okano H, Shiraki K, Inoue H, et al (2003) Cellular FLICE/caspase-8-inhibitory protein as a principal regulator of cell death and survival in human hepatocellular carcinoma. Lab Invest 83:1033–043
Perlman H, Pagliari LJ, Georganas C, Mano T, Walsh K, Pope RM (1999) FLICE-inhibitory protein expression during macrophage differentiation confers resistance to fas-mediated apoptosis. J Exp Med 190:1679–688
Tyler KL, Squier MK, Rodgers SE, et al (1995) Differences in the capacity of reovirus strains to induce apoptosis are determined by the viral attachment protein sigma 1. J Virol 69:6972–979
Clarke P, Meintzer SM, Spalding AC, Johnson GL, Tyler KL (2001) Caspase 8-dependent sensitization of cancer cells to TRAIL-induced apoptosis following reovirus-infection. Oncogene 20:6910–919
Slinker BK (1998) The statistics of synergism. J Mol Cell Cardiol 30:723–31
Sturzenbecker LJ, Nibert M, Furlong D, Fields BN (1987) Intracellular digestion of reovirus particles requires a low pH and is an essential step in the viral infectious cycle. J Virol 61:2351–361
Bortul R, Tazzari PL, Cappellini A, et al (2003) Constitutively active Akt1 protects HL60 leukemia cells from TRAIL-induced apoptosis through a mechanism involving NF-kappaB activation and cFLIP(L) up-regulation. Leukemia 17:379–89
Asakuma J, Sumitomo M, Asano T, Asano T, Hayakawa M (2003) Selective Akt inactivation and tumor necrosis actor-related apoptosis-inducing ligand sensitization of renal cancer cells by low concentrations of paclitaxel. Cancer Res 63: 1365–370
Ryu BK, Lee MG, Chi SG, Kim YW, Park JH (2001) Increased expression of cFLIP(L) in colonic adenocarcinoma. J Pathol 194:15–9
Jonsson G, Paulie S, Grandien A (2003) High level of cFLIP correlates with resistance to death receptor-induced apoptosis in bladder carcinoma cells. Anticancer Res 23:1213–218
Griffith TS, Chin WA, Jackson GC, Lynch DH, Kubin MZ (1998) Intracellular regulation of TRAIL-induced apoptosis in human melanoma cells. J Immunol 161:2833–840
Brooks AD, Sayers TJ (2005) Reduction of the antiapoptotic protein cFLIP enhances the susceptibility of human renal cancer cells to TRAIL apoptosis. Cancer Immunol Immunother 54:499–05
Van Valen F, Fulda S, Schafer KL, et al (2003) Selective and nonselective toxicity of TRAIL/Apo2L combined with chemotherapy in human bone tumour cells vs. normal human cells. Int J Cancer 107:929–40
Shankar S, Singh TR, Fandy TE, Luetrakul T, Ross DD, Srivastava RK (2005) Interactive effects of histone deacetylase inhibitors and TRAIL on apoptosis in human leukemia cells: Involvement of both death receptor and mitochondrial pathways. Int J Mol Med 16:1125–138
El Zawahry A, Lu P, White SJ, Voelkel-Johnson C (2006) in vitro efficacy of AdTRAIL gene therapy of bladder cancer is enhanced by trichostatin A-mediated restoration of CAR expression and downregulation of cFLIP and Bcl-XL. Cancer Gene Ther 13:281–89
Shishodia S, Aggarwal BB (2004) Guggulsterone inhibits NF-kappaB and IkappaBalpha kinase activation, suppresses expression of anti-apoptotic gene products, and enhances apoptosis. J Biol Chem 279:47148–7158
Piva R, Gianferretti P, Ciucci A, Taulli R, Belardo G, Santoro MG (2005) 15-Deoxy-delta 12,14-prostaglandin J2 induces apoptosis in human malignant B cells: An effect associated with inhibition of NF-kappa B activity and down-regulation of antiapoptotic proteins. Blood 105:1750–758
Boehrer S, Nowak D, Puccetti E, et al (2006) Prostate-apoptosis-response-gene-4 increases sensitivity to TRAIL-induced apoptosis. Leuk Res 30:597–05
DeBiasi RL, Clarke P, Meintzer S, et al (2003) Reovirus-induced alteration in expression of apoptosis and DNA repair genes with potential roles in viral pathogenesis. J Virol 77:8934–947
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Clarke, P., Tyler, K.L. Down-regulation of cFLIP following reovirus infection sensitizes human ovarian cancer cells to TRAIL-induced apoptosis. Apoptosis 12, 211–223 (2007). https://doi.org/10.1007/s10495-006-0528-4
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DOI: https://doi.org/10.1007/s10495-006-0528-4