Dicoumarol sensitizes renal cell carcinoma Caki cells to TRAIL-induced apoptosis through down-regulation of Bcl-2, Mcl-1 and c-FLIP in a NQO1-independent manner

Eun Jung Park; Kyoung-Jin Min; Kyeong Sook Choi; Taeg Kyu Kwon

doi:10.1016/j.yexcr.2014.01.009

Dicoumarol sensitizes renal cell carcinoma Caki cells to TRAIL-induced apoptosis through down-regulation of Bcl-2, Mcl-1 and c-FLIP in a NQO1-independent manner

Exp Cell Res. 2014 Apr 15;323(1):144-154. doi: 10.1016/j.yexcr.2014.01.009. Epub 2014 Jan 22.

Authors

Eun Jung Park¹, Kyoung-Jin Min¹, Kyeong Sook Choi², Taeg Kyu Kwon³

Affiliations

¹ Department of Immunology, School of Medicine, Keimyung University, 2800 Dalgubeoldaero, Dalseo-Gu, Daegu 704-701, South Korea.
² Department of Biomedical Sciences, Ajou University School of Medicine, 5 Woncheon-Dong, Paldal-Gu, Suwon 442-749, South Korea.
³ Department of Immunology, School of Medicine, Keimyung University, 2800 Dalgubeoldaero, Dalseo-Gu, Daegu 704-701, South Korea. Electronic address: kwontk@dsmc.or.kr.

PMID: 24462458
DOI: 10.1016/j.yexcr.2014.01.009

Abstract

In the study, we investigated the effect of dicoumarol, an anti-coagulant agent with the inhibitory activity of NAD(P)H quinone oxidoreductase 1 (NQO1), on TRAIL-induced apoptosis in renal cancer cell. Combined treatment with dicoumarol and TRAIL significantly induced apoptosis in various human renal carcinoma cells including Caki, ACHN, and A498, but not in normal human skin fibroblasts (HSF) and mouse kidney cells (TMCK-1). When we elucidated the relevance of NQO1 in dicoumarol plus TRAIL-mediated apoptosis, both ES936 (a NQO1 inhibitor) and knockdown of NQO1 with siRNA had no effect on TRAIL-mediated apoptosis, suggesting that the stimulating effect of dicoumarol on TRAIL-mediated apoptosis is independent of NQO1 activity. We found that dicoumarol transcriptionally down-regulated Bcl-2 expression via inhibition of NF-κB and CREB activity, whereas it down-regulated Mcl-1 and c-FLIP expression at the post-translational level. Overexpression of Bcl-2, Mcl-1, or c-FLIP overcame the dicoumarol plus TRAIL-induced apoptosis, indicating that down-regualtion of these anti-apoptotic proteins may critically contribute to the sensitizing effect of dicoumarol on TRAIL-mediated apoptosis.

Keywords: Bcl-2; Dicoumarol; Mcl-1; NQO1; TRAIL; c-FLIP.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apoptosis / drug effects
Apoptosis / physiology*
CASP8 and FADD-Like Apoptosis Regulating Protein / biosynthesis
CASP8 and FADD-Like Apoptosis Regulating Protein / metabolism
CREB-Binding Protein / antagonists & inhibitors
Carcinoma, Renal Cell / metabolism*
Cell Line, Tumor
Dicumarol / pharmacology*
Down-Regulation
Enzyme Inhibitors / pharmacology*
Humans
Indolequinones / pharmacology
Kidney Neoplasms / metabolism*
Mice
Myeloid Cell Leukemia Sequence 1 Protein / biosynthesis
Myeloid Cell Leukemia Sequence 1 Protein / metabolism
NAD(P)H Dehydrogenase (Quinone) / antagonists & inhibitors
NAD(P)H Dehydrogenase (Quinone) / genetics
NAD(P)H Dehydrogenase (Quinone) / metabolism
NF-kappa B / antagonists & inhibitors
Proto-Oncogene Proteins c-bcl-2 / biosynthesis
Proto-Oncogene Proteins c-bcl-2 / metabolism
RNA Interference
RNA, Small Interfering
Reactive Oxygen Species / metabolism
Signal Transduction / drug effects
TNF-Related Apoptosis-Inducing Ligand / metabolism*
Transcription Factor CHOP / genetics
Transcription, Genetic / drug effects

Substances

5-methoxy-1,2-dimethyl-3-((4-nitrophenoxy)methyl)indole-4,7-dione
CASP8 and FADD-Like Apoptosis Regulating Protein
CFLAR protein, human
DDIT3 protein, human
Enzyme Inhibitors
Indolequinones
MCL1 protein, human
Myeloid Cell Leukemia Sequence 1 Protein
NF-kappa B
Proto-Oncogene Proteins c-bcl-2
RNA, Small Interfering
Reactive Oxygen Species
TNF-Related Apoptosis-Inducing Ligand
TNFSF10 protein, human
Transcription Factor CHOP
Dicumarol
NAD(P)H Dehydrogenase (Quinone)
NQO1 protein, human
CREB-Binding Protein
CREBBP protein, human