Characterization of the aryl hydrocarbon receptor in the human C-4II cervical squamous carcinoma cell line

Abstract

Treatment of C-4II human cervical squamous carcinoma cells with 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD) gave a concentration-dependent increase in ethoxyresorufin O-deethylase (EROD) activity. The EC 50 for this response was approximately 1 nM and the maximum induced activity was 27 pmol/min/mg protein. The molecular properties of the cytosolic and nuclear aryl hydrocarbon (Ah) receptor complex were determined by velocity sedimentation analysis, photoaffinity labeling, gel retardation using a consensus dioxin responsive element (DRE), and DNA-Sepharose, DRE-Sepharose and Sephaeryl S-300 gel permeation column chromatography. The apparent molecular masses of the cytosolic and nuclear photoaffinity-labeled Ah receptor complexes were 110kDa and differed from the corresponding values obtained for the Ah receptor from other animal species. In contrast, most of the other molecular properties of the Ah receptor were not significantly different from those previously reported for other species. The relative Ah-responsiveness of the C-4II cells was assessed by determining the ratio of the induced EROD activity/nuclear Ah receptor levels for a submaximal inducing dose of [ 3H]TCDD. The induced activity/binding ratio for the human C-4II cells was 0.77 and was at least one order of magnitude lower than the corresponding value for the Ah-responsive rat hepatoma H-4-II E cells.