Supplemental Material
Estrogenicity and androgenicity screening of PCB sulfate monoesters in human breast cancer MCF-7 cells
Susanne Flor, Xianran He, Hans-Joachim Lehmler, Gabriele Ludewig
Department of Occupational & Environmental Health, College of Public Health, The University of Iowa, Iowa City, IA
Figure S1: Cytotoxicity of PCB sulfates and 4’-HO-PCB 3 in MCF7-BOS cells after 120 h exposure. The concentrations tested were from left to right: (A) 0.005, 0.05, 1.25, 5, 12.5, 25, 50 and 100 µM and (B) 0.01 and 1nM, 0.01, 1 and 100 µM. The percent of viable cells was determined using the resazurin assay. Only 4’-HO-PCB 3 was 100% toxic at 100 µM, 4PCB 53 sulfate was weakly cytotoxic (84% viability).
Fig. S2-1: Estrogenic Proliferative Effect of four PCB sulfates.
PCB sulfate proliferative data are normalized against the control (0.5% DMSO). Data depicted are the mean of 2 individual experiments with each duplicate samples ± SE. Levels of significance: * p < 0.05; ** p < 0.01 and *** p < 0.001.
Fig. S2-2: Anti-Estrogenic Effect of three PCB sulfates and 4’-HO-PCB 3.
Proliferative data of test compounds are normalized to the control (100 pM E2). Data depicted are the mean of 2 individual experiments with each duplicate samples ± SE. Levels of significance: * p < 0.05; ** p < 0.01 and *** p < 0.001.
Fig. S3-1: Androgenic Effect of three PCB sulfates.
Proliferative data of test compounds are normalized to the control (100 pM E2). Data depicted are the mean of 2 individual experiments with duplicate samples ± SE. Levels of significance: * p < 0.05; ** p < 0.01 and *** p < 0.001
Fig. S3-2: Anti-Androgenic Proliferative Effect of seven PCB sulfates and 4’-HO-PCB 3. Test compound data are normalized to the control (100 pM E2 + 500 pM R1881). Data depicted are the mean of 2 individual experiments with duplicate samples ± SE. Levels of significance: * p < 0.05; ** p < 0.01 and *** p < 0.001
Fig. S4: Co-incubation with the anti-estrogen ICI 182,780 abolished the estrogenic effect of PCB sulfates and 4’-HO-PCB 3 in MCF7-BOS cells.
10 nM ICI 182,780 was added to estrogenic concentrations of 2’PCB 3, 4’PCB 3, 4PCB 39 and 4PCB 53 sulfate (100 µM) and 4’-HO-PCB 3 (1 µM). ICI 182,780 completely inhibited the induction of proliferation by the PCB metabolites. Data depicted are the mean of 2 individual experiments in duplicates ± SE. Levels of significance are indicated as: * p < 0.05; ** p < 0.01 and *** p < 0.001
Fig. S5: Example of standard curves for E2 exposure in MCF7-BOS cells and R1881 with E2 (100 pM) co-exposure in MCF7-AR1 cells.
Controls and standard curves were routinely included in each individual experiment and evaluated for consistence with historical data.