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Death induction by recombinant native TRAIL and its prevention by a caspase 9 inhibitor in primary human esophageal epithelial cells

 Seok-Hyun Kim  ;  Kunhong Kim  ;  Wafik S. El-Deiry  ;  Youhai Chen  ;  Anil K. Rustgi  ;  Meenhard Herlyn  ;  David T. Dicker  ;  Jae G. Kwagh 
 JOURNAL OF BIOLOGICAL CHEMISTRY, Vol.279(38) : 40044-40052, 2004 
Journal Title
Issue Date
Adenocarcinoma ; Antineoplastic Agents/pharmacology* ; Caspase 9 ; Caspase Inhibitors* ; Caspases/metabolism ; Cell Death/drug effects ; Cell Line, Tumor/drug effects ; Epithelial Cells/cytology ; Epithelial Cells/drug effects* ; Esophagus/cytology ; GPI-Linked Proteins ; Humans ; Lung Neoplasms ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; Receptors, Tumor Necrosis Factor/genetics* ; Receptors, Tumor Necrosis Factor/therapeutic use ; Receptors, Tumor Necrosis Factor, Member 10c ; Recombinant Proteins/genetics ; Recombinant Proteins/toxicity* ; Tumor Necrosis Factor Decoy Receptors
The cytotoxic death ligand TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) is a tumor-specific agent under development as a novel anticancer therapeutic agent. However, some reports have demonstrated toxicity of certain TRAIL preparations toward human hepatocytes and keratinocytes through a caspase-dependent mechanism that involves activation of the extrinsic death pathway and Type II signaling through the mitochondria. We have isolated and purified both His-tagged protein and three versions of native recombinant human TRAIL protein from Escherichia coli. We found that 5 mm dithiothreitol in the purification process enhanced oligomerization of TRAIL and resulted in the formation of hyper-oligomerized TRAILs, including hexamers and nonomers with an extremely high potency in apoptosis induction. Although death-inducing signaling complex formation was much more efficient in cells treated with hyper-oligomerized TRAILs, this did not convert TRAIL-sensitive Type II HCT116 colon tumor cells to a Type I death pattern as judged by their continued sensitivity to a caspase 9 inhibitor. Moreover, TRAIL-resistant Type II Bax-null colon carcinoma cells were not converted to a TRAIL-sensitive Type I state by hyper-oligomerized TRAIL. Primary human esophageal epithelial 2 cells were found to be sensitive to all TRAIL preparations used, including trimer TRAIL. TRAIL-induced death in esophageal epithelial 2 cells was prevented by caspase 9 inhibition for up to 4 h after TRAIL exposure. This result suggests a possible therapeutic application of caspase 9 inhibition as a strategy to reverse TRAIL toxicity. Hyper-oligomerized TRAIL may be considered as an alternative agent for testing in clinical trials.
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1. College of Medicine (의과대학) > Dept. of Biochemistry and Molecular Biology (생화학-분자생물학교실) > 1. Journal Papers
Yonsei Authors
Kim, Kun Hong(김건홍) ORCID logo https://orcid.org/0000-0001-5639-6372
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