Cell-Specific Regulation of Human Aryl Hydrocarbon Receptor Expression by Transforming Growth Factor-β1
- Sandra Wolff1,
- Patricia A. Harper2,
- Judy M. Y. Wong2,
- Volker Mostert1,
- Yanping Wang2 and
- Josef Abel1
- 1Department of Experimental Toxicology, Medical Institute of Environmental Hygiene at the Heinrich-Heine-University, Düsseldorf, Germany (S.W., V.M., J.A.); and 2Division of Clinical Pharmacology, Hospital for Sick Children, Toronto, Ontario, Canada (P.A.H., J.M.Y.W., Y.W.)
Abstract
Previous studies showed that TGF-β down-regulates aryl hydrocarbon (AhR) expression in human lung carcinoma cells A549. Here we analyzed the molecular mechanisms by which TGF-β modulates AhR expression. A 5799-nucleotide 5′-flanking region of human AhR gene was isolated. Transient transfection studies of full-length (hAhRP) and deletion promoter constructs indicate the requirement of acis-regulatory element encompassing −1980 to −1892 for full constitutive activity. Basal hAhRP activity occurs in a cell-specific manner; human hepatoma HepG2 cells possess a 10-fold higher activity compared with A549 cells. TGF-β exerts cell-specific effects on hAhRP activity. Treatment of cells with 100 pM TGF-β leads to a 50% inhibition in A549 and a 3-fold induction in HepG2 cells. Deletion mutagenesis identified a TGF-β-responsive sequence containing a functional conserved Smad-binding element. Transient overexpression of Smad 2, 3, and 4 indicates that these signal transducers modulate hAhRP activity. The down-regulation of AhR by TGF-β is modulated by 5′-TG-3′-interacting factor (TGIF). Transient overexpression of TGIF in MDA-MB231 and HepG2 cells led to inhibition of hAhRP activity and a similar decrease of AhR mRNA expression. Our findings indicate that Smad proteins are involved in the cell-specific regulation of AhR expression by TGF-β.
Footnotes
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Send reprint requests to: Dr. Joseph Abel, Medical Institute of Environmental Hygiene at the Heinrich-Heine-University Düsseldorf, Department of Experimental Toxicology, Auf'm Hennekamp 50, 40225 Düsseldorf, Germany. E-mail:josef.abel{at}uni-duesseldorf.de
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This work was supported by the Deutsche Forschungsgemeinschaft, Sonderforschungsbereich 503/A5 and by a grant from the National Cancer Institute of Canada to P.A.H.
- Abbreviations:
- AhR
- aryl hydrocarbon receptor
- bHLH
- basic helix-loop-helix
- PAS
- Per-Arnt-Sim (periodicity/aryl hydrocarbon receptor nuclear translocator/simple-minded)
- Arnt
- aryl hydrocarbon receptor nuclear translocator
- TCDD
- 2,3,7,8-tetrachlorodibenzo-p-dioxin
- SBE
- Smad binding element
- TGIF
- 5′-TG-3′-interacting factor
- BMS
- basal medium supplement
- FCS
- fetal calf serum
- nt
- nucleotides
- AhRP
- aryl hydrocarbon receptor promoter
- kb
- kilobase pair(s)
- SBS
- Smad binding sequence
- SBCE
- Smad binding core element
- EMSA
- electrophoretic mobility shift assay
- DMEM
- Dulbecco's modified Eagle's medium
- PCR
- polymerase chain reaction
- TGF-β
- transforming growth factor-β
- TβR
- transforming growth factor-β receptor
- RT
- reverse transcription
- Cdk
- cyclin-dependent kinase
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- Received August 17, 2000.
- Accepted December 20, 2000.
- The American Society for Pharmacology and Experimental Therapeutics
