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. Author manuscript; available in PMC: 2015 May 1.

Published in final edited form as: Cancer Prev Res (Phila). 2014 Mar 5;7(5):505–515. doi: 10.1158/1940-6207.CAPR-13-0296

(A) A significant induction in SULT 1 enzyme expression was observed by immunoblotting on co-treatment of MCF-10A cells with DMA or Ral. Relative protein amounts were determined by densitometric analysis of SULT1 protein after western blot, loading with 30 μg of total protein. Each treatment was normalized to the loading and transferring control, β actin. Each data point represents an average of three independent experiments in duplicate ± SD; * p < 0.05. (B) Gene transcription of SULT 1E1 was significantly induced by DMA and Ral while FDMA had no effect compare to E₂ treatment alone. (C) Gene transcription of UGT 1A1 was significantly induced co-treatment with DMA or Ral as measured by qPCR after isolating RNA from 24 h treated MCF-10A cells. Each data point represents an average of three independent experiments ± SD; * p < 0.05, ** p < 0.01. (D) Co-treatment with DMA or Ral significantly inhibited E₂-induced anchorage-independent colony growth of MCF-10A cells in soft agar, while FDMA co-treatment had little effect. Cells were treated twice a week, in the presence and absence of SERMs, over the course of 3 weeks. DMSO (0.01%) was used as the vehicle control in the experiments in the absence of E₂ treatment. Cells were plated on soft agar and maintained for 3 weeks. Relative colony efficiency is calculated by dividing the number of colonies counted in a well by the number of cells plated in each well, normalized to DMSO vehicle. Data shows mean from three independent experiments ± SD: ** p < 0.005.

(A) A significant induction in SULT 1 enzyme expression was observed by immunoblotting on co-treatment of MCF-10A cells with DMA or Ral. Relative protein amounts were determined by densitometric analysis of SULT1 protein after western blot, loading with 30 μg of total protein. Each treatment was normalized to the loading and transferring control, β actin. Each data point represents an average of three independent experiments in duplicate ± SD; * p < 0.05. (B) Gene transcription of SULT 1E1 was significantly induced by DMA and Ral while FDMA had no effect compare to E₂ treatment alone. (C) Gene transcription of UGT 1A1 was significantly induced co-treatment with DMA or Ral as measured by qPCR after isolating RNA from 24 h treated MCF-10A cells. Each data point represents an average of three independent experiments ± SD; * p < 0.05, ** p < 0.01. (D) Co-treatment with DMA or Ral significantly inhibited E₂-induced anchorage-independent colony growth of MCF-10A cells in soft agar, while FDMA co-treatment had little effect. Cells were treated twice a week, in the presence and absence of SERMs, over the course of 3 weeks. DMSO (0.01%) was used as the vehicle control in the experiments in the absence of E₂ treatment. Cells were plated on soft agar and maintained for 3 weeks. Relative colony efficiency is calculated by dividing the number of colonies counted in a well by the number of cells plated in each well, normalized to DMSO vehicle. Data shows mean from three independent experiments ± SD: ** p < 0.005.