Combination of IFNγ and Chemotherapeutic Agents Increase TRAIL Sensitivity of Neuroblastoma Cell Lines

Abstract

Objective: Aim of the study was to further evaluate the role of caspase 8 and death receptors (DR) in the TRAIL-induced apoptosis of neuroblastoma (NB) cell lines.

Methods: Caspase 8 mRNA expression was monitored by RT-PCR. Caspase 8, DR5 and DR4 protein expression were monitored by Western blot analysis. The effects of IFNγ, TRAIL, IFNγ+TRAIL, caspase 8 inhibitor+TRAIL and IFNγ+chemotherapeutic agents+TRAIL on the growth and apoptosis of NB cells were detected with MTT and flow cytometry. The relative caspase 8 activity was measured with colorimetric assay.

Results: Caspase 8 expression was induced by IFNγ in the NB cell line SKNDZ. TRAIL alone did not induce apoptosis compared with controls but a combination of IFNγ+TRAIL and IFNγ+chemotherapeutic agents+TRAIL significantly induced cell apoptosis in SY5Y cells. Etoposide and doxorubicin induced DR5 but not DR4 in NB cell lines. SKNDZ cells expressing caspase 8 after treatment with IFNγ were still resistant to TRAIL but sensitive to TRAIL after the induction of DR5.

Conclusions: Sensitization of NB cells to TRAIL may be mediated by the upregulation of caspase 8 with IFNγ and DR5 with chemotherapeutic agents. This suggests that caspase 8 and death receptors play a very important role in TRAIL-induced apoptosis of NB cells and a combination of IFNγ, TRAIL and chemotherapeutic agents may be a new and interesting anticancer treatment strategy for NB.

neuroblastoma - caspase 8 - TRAIL - death receptor

References

• 1 Fulda S, Debatin KM. IFN gamma sensitizes for apoptosis by upregulating caspase-8 expression through the Stat1 pathway.  Oncogene. 2002;  21 2295-2308
• 2 Yang X, Merchant MS, Romero ME. et al . Induction of caspase 8 by interferon gamma renders some neuroblastoma cells sensitive to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) but reveals that a lack of membrane TR₁/TR₂ also contributes to TRAIL resistance in neuroblastoma.  Cancer Res. 2003;  63 1122-1129
• 3 Fulda S, Debatin KM. Modulation of TRAIL signaling for cancer therapy.  Vitam Horm. 2004;  67 275-290
• 4 Tong HX, Lu CW, Yang YM. et al . Role of caspase 8 as a determinant in TRAIL sensitivity of neuroblastoma cell lines.  Pediatr Hemat Oncol. 2009;  26 549-559
• 5 Tong HX, Lu CW, Zhang JH. et al . Enhanced effect of IFNγ on the induced-apoptosis of neuroblastoma cells by cytotoxic drugs.  Pediatr Hemat Oncol. 2008;  25 549-558
• 6 Tong HX, Lu CW, Zhang JH. et al . Combination of γ-Interferon with TRAIL and cisplatin or etoposide induces apoptosis in neuroblastoma cell line SH-SY5Y.  Chin Med Sci J. 2007;  22 38-43
• 7 Ebinger M, Senf L, Wachowski O. et al . Promoter methylation pattern of caspase-8, P16INK4A, MGMT, TIMP-3, and E-cadherin in medulloblastoma.  Pathol Oncol Res. 2004;  10 17-21
• 8 Fulda S, Debatin KM. 5-Aza-2′-deoxycytidine and IFN-gamma cooperate to sensitize for TRAIL-induced apoptosis by upregulating caspase-8.  Oncogene. 2006;  25 5125-5133
• 9 Tekautz TM, Zhu K, Grenet J. et al . Evaluation of IFN-gamma effects on apoptosis and gene expression in neuroblastoma – preclinical studies.  Biochim Biophys Acta. 2006;  1763 1000-1010
• 10 Stupack DG, Teitz T, Potter MD. et al . Potentiation of neuroblastoma metastasis by loss of caspase-8.  Nature. 2006;  439 95-99
• 11 McKee AE, Thiele CJ. Targeting caspase 8 to reduce the formation of metastases in neuroblastoma.  Expert Opin Ther Targets. 2006;  10 703-708
• 12 Wang MJ, Liu YX, Li XL. et al . Chemotherapeutic drugs enhanced rsTRAIL tumoricidal activity.  Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2004;  26 524-528
• 13 Kim YH, Lee YJ. Time sequence of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and cisplatin treatment is responsible for a complex pattern of synergistic cytotoxicity.  J Cell Biochem. 2006;  98 1284-1295
• 14 Day TW, Wu CH, Safa AR. Etoposide induces protein kinase Cdelta- and caspase-3-dependent apoptosis in neuroblastoma cancer cells.  Mol Pharmacol. 2009;  76 632-640
• 15 Frese S, Frese-Schaper M, Andres AC. et al . Cardiac glycosides initiate Apo2L/TRAIL-induced apoptosis in non-small cell lung cancer cells by up-regulation of death receptors 4 and 5.  Cancer Res. 2006;  66 5867-5874
• 16 Karacay B, Sanlioglu S, Griffith TS. et al . Inhibition of the NF-kappa-B pathway enhances TRAIL-mediated apoptosis in neuroblastoma cells.  Cancer Gene Ther. 2004;  11 681-690
• 17 van der Sloot AM, Tur V, Szegezdi E. et al . Designed tumor necrosis factor-related apoptosis-inducing ligand variants initiating apoptosis exclusively via the DR5 receptor.  Proc Natl Acad Sci USA. 2006;  103 8634-8639
• 18 Jin H, Yang R, Fong S. et al . Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand cooperates with chemotherapy to inhibit orthotopic lung tumor growth and improve survival.  Cancer Res. 2004;  64 4900-4905
• 19 Liu P, Mao H, Hou P. Synergistic antitumor effect of tumor necrosis factor-related apoptosis-inducing ligand combined with cisplatin in ovarian carcinoma cell lines in vitro and in vivo.  Int J Gynecol Cancer. 2006;  16 538-548
• 20 Song K, Benhaga N, Anderson RL. et al . Transduction of tumor necrosis factor-related apoptosis-inducing ligand into hematopoietic cells leads to inhibition of syngeneic tumor growth in vivo.  Cancer Res. 2006;  66 6304-6311

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