Abstract | SCOPE: METHODS AND RESULTS: HepG2 cells were treated with 6-MTITC with varying time and dose. NQO1, Nrf2, and Keap1 proteins were detected by Western blotting. ARE transactivation was detected by electrophilic mobility shift assay and reporter gene assay. Nuclear localization of Nrf2 was determined by immunocytochemistry assay. Ubiquitination of Nrf2 and Keap1 was detected using immunoprecipitation after treatment with MG132. Small interfering RNA was used to knockdown Nrf2 or Keap1. The results revealed that 6-MTITC modulated Nrf2/ARE pathway by stimulating Keap1 modification, and inhibiting Nrf2 ubiquitination and protein turnover. These actions finally increased nuclear Nrf2 accumulation and ARE-binding activity. Moreover, silencing Nrf2 markedly reduced ARE-driven activity induced by 6-MTITC. CONCLUSION:
6-MTITC modulated ARE-driven NQO1 expression by stabilizing Nrf2 with enhanced Keap1 modification and decreased Nrf2 degradation.
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Authors | Yoshimi Korenori, Shunsuke Tanigawa, Takuma Kumamoto, Si Qin, Yosuke Daikoku, Koji Miyamori, Masashi Nagai, De-Xing Hou |
Journal | Molecular nutrition & food research
(Mol Nutr Food Res)
Vol. 57
Issue 5
Pg. 854-64
(May 2013)
ISSN: 1613-4133 [Electronic] Germany |
PMID | 23390006
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Copyright | © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. |
Chemical References |
- Antioxidants
- Intracellular Signaling Peptides and Proteins
- Isothiocyanates
- KEAP1 protein, human
- Kelch-Like ECH-Associated Protein 1
- NF-E2-Related Factor 2
- NFE2L2 protein, human
- 6-methylthiohexyl isothiocyanate
- NAD(P)H Dehydrogenase (Quinone)
- NQO1 protein, human
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Topics |
- Antioxidants
(pharmacology)
- Cytoprotection
- Electrophoretic Mobility Shift Assay
- Gene Expression Regulation
- Genes, Reporter
- Hep G2 Cells
- Humans
- Intracellular Signaling Peptides and Proteins
(genetics, metabolism)
- Isothiocyanates
(pharmacology)
- Kelch-Like ECH-Associated Protein 1
- NAD(P)H Dehydrogenase (Quinone)
(genetics, metabolism)
- NF-E2-Related Factor 2
(genetics, metabolism)
- Response Elements
(drug effects)
- Signal Transduction
- Wasabia
(chemistry)
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