Abstract
Because Bcl-2 family members inhibit the ability of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) to induce apoptosis, we investigated whether ABT-737, a small molecule Bcl-2 inhibitor, enhances TRAIL killing. We demonstrate that a combination of ABT-737 and TRAIL induced significant cell death in multiple cancer types, including renal, prostate, and lung cancers, although each agent individually had little activity in these tumor cells. All of these cell lines expressed the Mcl-1 protein that is known to block the activity of ABT-737 and TRAIL but did not block the synergy between these agents. However, Bax-deficient cell lines, including DU145 and HCT116 cells and those cell lines expressing low levels of TRAIL receptor, were resistant to apoptosis induced by these agents. To understand how ABT-737 functions to markedly increase TRAIL sensitivity, the levels of specific death-inducing signaling complex components were evaluated. Treatment with ABT-737 did not change the levels of c-FLIP, FADD, and caspase-8 but up-regulated the levels of the TRAIL receptor DR5. DR5 up-regulation induced by ABT-737 treatment occurred through a transcriptional mechanism, and mutagenesis studies demonstrated that the NF-kappaB site found in the DR5 promoter was essential for the ability of ABT-737 to increase the levels of this mRNA. Using luciferase reporter plasmids, ABT-737 was shown to stimulate NF-kappaB activity. Together, these results demonstrate that the ability of ABT-737 and TRAIL to induce apoptosis is mediated through activation of both the extrinsic and intrinsic pathways. Combinations of ABT-737 and TRAIL can be exploited therapeutically where antiapoptotic Bcl-2 family members drive tumor cell resistance to current anticancer therapies.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Antineoplastic Agents / pharmacology*
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Antineoplastic Agents / therapeutic use
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Apoptosis / drug effects*
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Apoptosis / genetics
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Biphenyl Compounds / pharmacology*
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Biphenyl Compounds / therapeutic use
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CASP8 and FADD-Like Apoptosis Regulating Protein / genetics
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CASP8 and FADD-Like Apoptosis Regulating Protein / metabolism
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Caspase 8 / genetics
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Caspase 8 / metabolism
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Cell Line, Tumor
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Drug Screening Assays, Antitumor / methods
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Fas-Associated Death Domain Protein / genetics
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Fas-Associated Death Domain Protein / metabolism
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Humans
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Mutagenesis
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Myeloid Cell Leukemia Sequence 1 Protein
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NF-kappa B / genetics
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NF-kappa B / metabolism
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Neoplasms / drug therapy*
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Neoplasms / metabolism
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Nitrophenols / pharmacology*
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Nitrophenols / therapeutic use
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Piperazines / pharmacology
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Piperazines / therapeutic use
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Proto-Oncogene Proteins c-bcl-2 / antagonists & inhibitors*
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Proto-Oncogene Proteins c-bcl-2 / genetics
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Proto-Oncogene Proteins c-bcl-2 / metabolism
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Receptors, TNF-Related Apoptosis-Inducing Ligand / biosynthesis*
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Receptors, TNF-Related Apoptosis-Inducing Ligand / genetics
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Response Elements / genetics
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Sulfonamides / pharmacology*
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Sulfonamides / therapeutic use
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TNF-Related Apoptosis-Inducing Ligand / pharmacology*
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Up-Regulation / drug effects
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Up-Regulation / genetics
Substances
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ABT-737
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Antineoplastic Agents
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Biphenyl Compounds
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CASP8 and FADD-Like Apoptosis Regulating Protein
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CFLAR protein, human
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FADD protein, human
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Fas-Associated Death Domain Protein
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Myeloid Cell Leukemia Sequence 1 Protein
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NF-kappa B
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Nitrophenols
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Piperazines
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Proto-Oncogene Proteins c-bcl-2
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Receptors, TNF-Related Apoptosis-Inducing Ligand
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Sulfonamides
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TNF-Related Apoptosis-Inducing Ligand
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TNFSF10 protein, human
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CASP8 protein, human
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Caspase 8