Abstract
TNF-related apoptosis-inducing ligand (TRAIL), a novel member of the tumor necrosis factor (TNF) family, is thought to induce apoptosis preferentially in cancer cells; however, increasing evidence suggests that a number of cancers are resistant to TRAIL treatment. FLICE-like inhibitory protein (FLIP), which structurally resembles caspase-8, can act as an inhibitor of apoptosis when expressed at high levels in certain cancer cells. The purpose of our present study was to determine whether human colon cancer cells are sensitive to TRAIL treatment and, if not, to identify potential mechanisms of resistance. Colon cancer cells of different metastatic potential (KM12C, KML4A, and KM20) were found to be resistant to the effects of TRAIL when used as a single agent. FLIP expression levels were increased in all three KM cell lines. Treatment with either actinomycin D (Act D;10 :g/ml) or cycloheximide (CHX; 10 :g/ml) decreased FLIP expression levels in all three cell lines. The decrease in cellular levels of FLIP was associated with sensitization to TRAIL-mediated apoptosis, as demonstrated by enhanced cell death and caspase-3 activity compared with either Act D or CHX alone. Our findings suggest that reduction of FLIP levels by Act D or CHX renders TRAIL-resistant human colon cancer cells sensitive to TRAIL-mediated apoptosis. The combination of TRAIL along with agents such as Act D or CHX, which target proteins that prevent cell death, may provide a more effective and less toxic regimen for treatment of resistant colon cancers.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Antibiotics, Antineoplastic / pharmacology
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Antibiotics, Antineoplastic / therapeutic use*
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Antineoplastic Combined Chemotherapy Protocols / pharmacology
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Antineoplastic Combined Chemotherapy Protocols / therapeutic use*
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Apoptosis / drug effects*
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Apoptosis / physiology*
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Apoptosis Regulatory Proteins
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Blotting, Western
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Caspase 8
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Caspase 9
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Caspases / analysis*
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Caspases / drug effects*
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Caspases / physiology
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Colonic Neoplasms
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Cycloheximide / pharmacology
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Cycloheximide / therapeutic use*
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Dactinomycin / pharmacology
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Dactinomycin / therapeutic use*
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Drug Evaluation, Preclinical
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Drug Resistance, Neoplasm
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Gene Expression Regulation, Neoplastic / drug effects*
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Gene Expression Regulation, Neoplastic / physiology*
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Humans
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Liver Neoplasms / secondary
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Membrane Glycoproteins / drug effects*
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Membrane Glycoproteins / pharmacology
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Membrane Glycoproteins / therapeutic use*
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Protein Synthesis Inhibitors / pharmacology
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Protein Synthesis Inhibitors / therapeutic use*
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RNA / analysis
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RNA / genetics
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Reverse Transcriptase Polymerase Chain Reaction
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TNF-Related Apoptosis-Inducing Ligand
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Tumor Cells, Cultured / drug effects
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Tumor Cells, Cultured / physiology
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Tumor Necrosis Factor-alpha / drug effects*
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Tumor Necrosis Factor-alpha / pharmacology
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Tumor Necrosis Factor-alpha / therapeutic use*
Substances
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Antibiotics, Antineoplastic
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Apoptosis Regulatory Proteins
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Membrane Glycoproteins
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Protein Synthesis Inhibitors
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TNF-Related Apoptosis-Inducing Ligand
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TNFSF10 protein, human
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Tumor Necrosis Factor-alpha
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Dactinomycin
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RNA
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Cycloheximide
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CASP8 protein, human
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CASP9 protein, human
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Caspase 8
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Caspase 9
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Caspases