Transcriptional regulation of human carboxylesterase 1A1 by nuclear factor-erythroid 2 related factor 2 (Nrf2).
Human carboxylesterase (CES) 1A, which is predominantly expressed in liver and lung, plays an important role in the hydrolysis of endogenous compounds and xenobiotics. CES1A is reported to be induced in human hepatocytes by butylated hydroxyanisole, ticlopidine and diclofenac, and the induction is assumed to be caused by oxidative stress. However, the molecular mechanism remains to be determined. In this study, we sought to investigate whether CES1A is regulated by nuclear factor-erythroid 2 related factor 2 (Nrf2), which is a transcriptional factor activated by oxidative stress, and clarify the molecular mechanism. Real-time reverse transcription-PCR assays revealed that CES1A1 mRNA was significantly induced by tert-butylhydroquinone (tBHQ) and sulforaphane (SFN), which are representative activators of Nrf2 in HepG2, Caco-2 and HeLa cells. The induction was completely suppressed with small interfering RNA for Nrf2. In HepG2 cells, the CES1A protein level and imidapril hydrolase activity, which is specifically catalyzed by CES1A, were also significantly induced by tBHQ and SFN. Luciferase assays revealed that the antioxidant response element (ARE) at -2025 in the CES1A1 gene was responsible for the transactivation by Nrf2. In addition, electrophoretic mobility shift assays and chromatin immunoprecipitation assays revealed that Nrf2 binds to the ARE in the CES1A1 gene. These findings clearly demonstrated that human CES1A1 is induced by Nrf2. This is the first study to demonstrate the molecular mechanism of the inducible regulation of human CES1A1.
Maruichi
Taiga
T
Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan.
Fukami
Tatsuki
T
Nakajima
Miki
M
Yokoi
Tsuyoshi
T
D001483:Base Sequence; D015153:Blotting, Western; D043182:Carboxylesterase / Q000235:genetics*; D002460:Cell Line; D017931:DNA Primers; D024202:Electrophoretic Mobility Shift Assay; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D006801:Humans; D051267:NF-E2-Related Factor 2 / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic / Q000502:physiology*; D002265:Carboxylic Ester Hydrolases
D043182:Carboxylesterase; D017931:DNA Primers; D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; C474609:CES1 protein, human; D002265:Carboxylic Ester Hydrolases