Figure 3.

The regulation of AR in the cell cycle of resistant cells. A. Having been treated with 100 nM DHT for 24, 48 or 72 hours, Cell viability of MCF-7 and MCF-7pR was measured by the Cell Counting kit-8 assay. *P < 0.05, or **P < 0.01 or ***P < 0.001. B. The expression of AR and CCND1 was detected by Western blot following the treatment of vehicle or 100 nM DHT for 48 hours in MCF-7 and MCF-7pR cells. C. The lysates of MCF-7pR treated with vehicle or 100 nM DHT were subjected to immunoprecipitation with AR antibodies. The immunoprecipitates were then blotted with indicated antibodies. D. AR mRNA levels were determined by real-time PCR in MCF-7pR cells following transfection with shAR or shNC for 48 hours. Data shown were from triplicate experiments and reported as the mean ± SD, ***P < 0.001. E. The expression of AR was detected by Western blot following transfection with shAR or shNC for 48 hours. *P < 0.05, or **P < 0.01 or ***P < 0.001. F. MCF-7pR cells were treated with DHT and/or transfected with shAR for 48 hours and measured by a flow cytometer. Results are reported as mean percent cell cycle distribution with standard errors. G. The protein expression of AR, CCND1, CDK2 and CCNE1 was determined by Western blot in MCF-7pR following treatment DHT and/or transfection with AR for 48 hours.