TCDD-dependent downregulation of gamma-catenin in rat liver epithelial cells (WB-F344).
TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) is the most potent tumor promoter ever tested in rodents. Although it is known that most of the effects of TCDD are mediated by binding to the aryl hydrocarbon receptor (AHR), the mechanisms leading to tumor promotion still remain to be elucidated. Loss of contact-inhibition is a characteristic hallmark in tumorigenesis. In WB-F344 cells, TCDD induces a release from contact-inhibition manifested by a 2- to 3-fold increase in DNA-synthesis and the emergence of foci when TCDD (1 nM) is given to confluent cells. We focussed our interest on potential cell membrane proteins mediating contact-inhibition in WB-F344 cells, namely E-cadherin, alpha,- beta,- and gamma-catenin (plakoglobin). Using indirect immunofluorescence, E-cadherin, alpha-, beta- and gamma-catenin were detected at cell adhesion sites in untreated, confluent cells. After TCDD-exposure, gamma-catenin was exclusively localized in the cytoplasm whereas localization of E-cadherin, alpha- and beta-catenin remained unaffected. Cytoplasmic gamma-catenin could be extracted by Triton X-100 treatment, demonstrating that gamma-catenin was no longer bound to the actin cytoskeleton. Western blot analysis showed downregulation of gamma-catenin protein levels. This effect was not blocked by pre-incubation with the selective proteasome inhibitor MG-132, indicating that proteolytical degradation of gamma-catenin by the proteasome system was not increased by TCDD. Because mRNA-levels of gamma-catenin were markedly diminished after TCDD-exposure, we conclude that transcriptional downregulation or destabilization of the mRNA contributes to the decrease in gamma-catenin protein levels in response to TCDD. Because gamma-catenin is considered to be a tumor suppressor, our findings might give more insight into the tumor promoting actions of TCDD.
Dietrich
Cornelia
C
Institute of Toxicology, Johannes Gutenberg-University, Obere Zahlbacherstrasse 67, 55131 Mainz, Germany. cdietric@mail.uni-mainz.de
Faust
Dagmar
D
Moskwa
Matthias
M
Kunz
Anja
A
Bock
Karl-Walter
KW
Oesch
Franz
F
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D015820:Cadherins / Q000378:metabolism; D002273:Carcinogens; D002455:Cell Division; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000096:biosynthesis* / Q000378:metabolism; D004247:DNA / Q000096:biosynthesis; D051180:Desmoplakins; D003902:Detergents / Q000494:pharmacology; D015536:Down-Regulation; D004785:Environmental Pollutants; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D008099:Liver / Q000166:cytology*; D008113:Liver Neoplasms / Q000473:pathology; D008856:Microscopy, Fluorescence; D009154:Mutation; D017830:Octoxynol / Q000494:pharmacology; D010641:Phenotype; D000072317:Polychlorinated Dibenzodioxins; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D015534:Trans-Activators / Q000378:metabolism; D051178:alpha Catenin; D051176:beta Catenin; D051185:gamma Catenin
D015820:Cadherins; D002273:Carcinogens; C495268:Ctnnb1 protein, rat; D003598:Cytoskeletal Proteins; D051180:Desmoplakins; D003902:Detergents; D004785:Environmental Pollutants; D000072317:Polychlorinated Dibenzodioxins; D015534:Trans-Activators; D051178:alpha Catenin; D051176:beta Catenin; D051185:gamma Catenin; D017830:Octoxynol; D004247:DNA