FIGURE 5.
The AR translational machinery is a target of 17-HASs. A and B, luciferase reporter assay, in which translation of the luciferase mRNA is under the control of AR 5′-UTR (test) or IRES (control). Cells were treated with 20 μm TOK-001 or an equal volume of vehicle (DMSO) in A or treated with increasing doses of TOK-001 in B. C and D, cap-binding assay with 7mG-pull down in total lysates from LNCaP cells treated with 10 μm or 20 μm TOK-001 in C or 10 μm or 20 μm abiraterone alcohol (ABI) in D for 36 h. Western blot analysis with the indicated antibodies is depicted. E, Western blot analysis of 4EPB1 and phospho-4EBP1 (p-4EBP1) in total lysates from LNCaP cells treated with vehicle (mock), 20 μm TOK-001, or 20 μm abiraterone alcohol (ABI). F, cell proliferation with MTS; *,** p < 0.007. LNCaP cells were incubated with mock (control), TOK-001 (10 μm), or abiraterone (10 μm) in serum-fed media for 72 h. Statistical analyses were performed using unpaired Student's t test. The data represent at least three independent experiments.