HDAC and mTORC1 inhibition in Myc-CaP cell lines in vitro result in alterations of androgen receptor and HIF-1α protein levels with loss of transcriptional activity.

(A) Myc-CaP cell lines with stable expression of androgen response element [Myc-ARE] plasmid treated with 10 nM R1881±10 nM panobinostat [P], 10 nM everolimus [E] or combination. At indicated times cells were lysed and luminescence intensity was measured and quantitated. Mean ± SE from 3 independent experiments. (B) AR and human c-Myc protein was investigated by immunoblot using whole cell Myc-CaP lysates after treatment with 10 nM R1881 with or without 10 nM panobinostat, 10 nM everolimus or combination for 24 hours. β-actin was used as loading control. (C) Myc-CaP cell lines with stable expression of hypoxia response element [Myc-HRE] plasmid treated with 100 µM CoCl2±10 nM panobinostat [P], 10 nM everolimus [E] or combination. At indicated times cells were lysed and luminescence intensity was measured and quantitated. Mean ± SE from 3 independent experiments. (D) HIF-1α protein was investigated by immunoblot using whole cell Myc-CaP lysates treated with 100 µM CoCl2 with or without 10 nM panobinostat, 10 nM everolimus or combination for 24 hours. β-actin was used as loading control. * p<0.05.