Glucocorticoid Receptor-Mediated Transcriptional Regulation of N-acetyltransferase 1 Gene Through Distal Promoter

Abstract

ABSTRACT. Human arylamine N-acetyltransferase 1, (HUMAN)NAT1, is a phase II xenobioticmetabolizing enzyme that plays an important role in drug and carcinogen biotransformation and cancer development. Its gene expression has been shown to be regulated by environmental factors. The purpose of the current study is to determine the involvement of nuclear receptors in transcriptional regulation of (HUMAN)NAT1 gene. We show that among the nuclear receptors examined, including the glucocorticoid receptor, retinoid acid receptor-related orphan receptor alpha, constitutive androstane receptor, pregnane X receptor, aryl hydrocarbon receptor, and retinoic acid receptor, the glucocorticoid receptor plays a dominant role in regulating (HUMAN)NAT1 gene expression through distal promoter (P3). The involvement of the glucocorticoid receptor in transcription regulation of (HUMAN)NAT1 gene expression was demonstrated by dexamethasone treatment, reporter assay using plasmid-containing 3 kbp of 5′-end region of promoter 3, and treatment of anti-glucocorticoid RU486 in primary culture of human hepatocytes and transfected HepG2 cells. In addition, translation inhibition did not affect dexamethasone-induced gene expression through P3, suggesting that dexamethasone effect is directly mediated by glucocorticoid receptor activation. Furthermore, deletion analysis revealed the presence of multiple responsive elements within the 3 kbp fragment of P3. Transfection assays in mice using hydrodynamics-based procedure and reporter gene assay in a mouse cell line revealed that glucocorticoid-induced NAT gene expression is species dependent. Dexamethasone treatment of transfected mice and mouse cell line decreased (MOUSE)Nat2 gene expression, (HUMAN)NAT1 homologue. These results suggest that glucocorticoids serve as a modulator for (HUMAN)NAT1 gene expression via the P3-containing 5′-flanking region.

Genetic and small molecule inhibition of arylamine N‐acetyltransferase 1 reduces anchorage‐independent growth in human breast cancer cell line MDA‐MB‐231

“…Human NAT1 activity is modified by polymorphism, but can also be regulated by microRNA, epigenetic, and/or translational and post‐translational control . NAT1 transcript levels can be regulated by extracellular stimuli acting on glucocorticoid or androgen receptors . Exposure to NAT1 substrates can increase NAT1 degradation, which results in decreased NAT1 activity .…”

Congenic rats with higher arylamine N-acetyltransferase 2 activity exhibit greater carcinogen-induced mammary tumor susceptibility independent of carcinogen metabolism

“…NAT1 activity is modified by genetic polymorphism, but also can be regulated by microRNA, epigenetic, and/or translational and post-translational control features [ 9 – 12 ]. NAT1 transcript levels are regulated by extracellular stimuli acting on glucocorticoid receptor or androgen receptor [ 13 , 14 ]. Exposure to NAT1 substrates can increase NAT1 degradation [ 11 , 12 ].…”

Role of human arylamine n-acetyltransferase 1 in tumorigenesis and cancer biology.

“…Human NAT1 activity is modified by polymorphism, but can also be regulated by microRNA, epigenetic, and/or translational and post-translational control features [67][68][69][70]. NAT1 transcript levels can be regulated by extracellular stimuli acting on glucocorticoid or androgen receptors [71,72]. Exposure to NAT1 substrates can increase NAT1 degradation, which results in decreased NAT1 activity [69,70].…”