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. 2017 Mar 23;8(21):34971–34979. doi: 10.18632/oncotarget.16496

Figure 2. Treatments with 8β-VE2 lead to downregulation of AR and ARVs in androgen-deprived prostate cancer cells.

Figure 2

(A) mRNA expression of the AR was measured by quantitative RT-PCR. VCaP rev, VCaP, and VCaP AA cells were treated with DMSO, 5 μM 8β-VE2 and 25 μM 8β-VE2. AR expression was strongly reduced after treatment with 8β-VE2 in VCaP control and VCaP AA cells. In VCaP rev cells, upregulation of AR mRNA expression could be observed. The data represent the mean±s.d. of three independent experiments, which were performed in duplicate. * P<0.05, ** P<0.01, *** P<0.0001 compared with DMSO control. (B) A representative western blot is shown of VCaP rev, VCaP, and VCaP AA cells treated with DMSO, 5 μM 8β-VE2, or 25 μM 8β-VE2. Total protein was extracted, and immunoblots were probed with AR-specific or ERβ-specific antibodies. A reduction in protein expression of the AR was detected in VCaP and in VCaP AA cells after 8β-VE2 treatment. In 8β-VE2-treated VCaP AA cells, the expression of ARVs was remarkably downregulated. In contrast, VCaP rev cells with low initial expression showed upregulated AR expression. In all three cell populations, downregulation of ERβ protein was observed at high 8β-VE2 concentration. The data represent two independent experiments, which were performed in duplicate. α-Tubulin was used as a loading control.