Proteasome inhibitor MG132 induces death receptor 5 through CCAAT/enhancer-binding protein homologous protein

Tatsushi Yoshida; Takumi Shiraishi; Susumu Nakata; Mano Horinaka; Miki Wakada; Yoichi Mizutani; Tsuneharu Miki; Toshiyuki Sakai

doi:10.1158/0008-5472.CAN-05-0693

Proteasome inhibitor MG132 induces death receptor 5 through CCAAT/enhancer-binding protein homologous protein

Cancer Res. 2005 Jul 1;65(13):5662-7. doi: 10.1158/0008-5472.CAN-05-0693.

Authors

Tatsushi Yoshida¹, Takumi Shiraishi, Susumu Nakata, Mano Horinaka, Miki Wakada, Yoichi Mizutani, Tsuneharu Miki, Toshiyuki Sakai

Affiliation

¹ Department of Molecular-Targeting Cancer Prevention, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kawaramachi-Hirokoji, Kyoto, Japan.

PMID: 15994939
DOI: 10.1158/0008-5472.CAN-05-0693

Abstract

Combined treatment with a proteasome inhibitor and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising strategy for cancer therapy. Proteasome inhibitors induce the expression of death receptor 5 (DR5), a receptor for TRAIL, and sensitize cancer cells to TRAIL-induced apoptosis; however, the molecular mechanism of DR5 up-regulation has not been elucidated. In this study, we report that CCAAT/enhancer-binding protein homologous protein (CHOP) is a regulator of DR5 induction by proteasome inhibitor MG132. MG132 induced DR5 expression at a protein and mRNA level in prostate cancer DU145 cells. Furthermore, MG132 increased DR5 promoter activity. Using a series of deletion mutant plasmids containing DR5 promoters of various sizes, we found that MG132 stimulated the promoter activity via the region of -289 to -253. This region contained a CHOP-binding site. Site-directed mutation of the site abrogated the promoter activity enhanced by MG132. An electrophoretic mobility shift assay showed that CHOP directly bound to the MG132-responsive site on the DR5 promoter. Expression of the CHOP protein was increased with MG132 along with DR5 up-regulation. Furthermore, CHOP small interfering RNA attenuated the DR5 up-regulation due to MG132. These results indicate that the proteasome inhibitor MG132 induces DR5 expression through CHOP up-regulation.

Publication types

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

MeSH terms

Apoptosis / drug effects
Apoptosis / physiology
Apoptosis Regulatory Proteins
CCAAT-Enhancer-Binding Proteins / metabolism
CCAAT-Enhancer-Binding Proteins / physiology*
Cell Line, Tumor
Cysteine Proteinase Inhibitors / pharmacology*
Humans
Leupeptins / pharmacology*
Male
Membrane Glycoproteins / pharmacology
Mutagenesis, Site-Directed
Promoter Regions, Genetic / drug effects
Prostatic Neoplasms / drug therapy
Prostatic Neoplasms / metabolism*
Prostatic Neoplasms / pathology
Proteasome Inhibitors*
RNA, Messenger / biosynthesis
RNA, Messenger / genetics
Receptors, TNF-Related Apoptosis-Inducing Ligand
Receptors, Tumor Necrosis Factor / biosynthesis*
Receptors, Tumor Necrosis Factor / genetics
TNF-Related Apoptosis-Inducing Ligand
Transcription Factor CHOP
Transcription Factors / metabolism
Transcription Factors / physiology*
Tumor Necrosis Factor-alpha / pharmacology
Up-Regulation / drug effects
Up-Regulation / physiology

Substances

Apoptosis Regulatory Proteins
CCAAT-Enhancer-Binding Proteins
Cysteine Proteinase Inhibitors
DDIT3 protein, human
Leupeptins
Membrane Glycoproteins
Proteasome Inhibitors
RNA, Messenger
Receptors, TNF-Related Apoptosis-Inducing Ligand
Receptors, Tumor Necrosis Factor
TNF-Related Apoptosis-Inducing Ligand
TNFRSF10B protein, human
TNFSF10 protein, human
Transcription Factors
Tumor Necrosis Factor-alpha
Transcription Factor CHOP
benzyloxycarbonylleucyl-leucyl-leucine aldehyde