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. 2017 Mar 22;8(19):31815–31829. doi: 10.18632/oncotarget.16463

Figure 1. Pharmacological inhibition of USP7 affects prostate cancer cell proliferation.

Figure 1

(A) Immunoblot analysis of USP7, CCDC6, AR and ARV7 isoform in human LNCaP, 22RV1 and PC3 prostate cancer cell lines. Antitubulin is shown as loading control. (B) LNCaP cells were treated with vehicle or different concentrations of P5091, as indicated, and cells were counted at the indicated times, in the presence or absence of DHT (10 nM). (C) PC3 cells were treated with vehicle or different concentration of P5091, as indicated, and cells were counted at the indicated times. In B and C the values are the mean +/− SD of three independent experiments. (D) USP7 inhibitors P5091 shows dose-dependent cytotoxic effect in prostate cancer cell lines. Cells were seeded in 96-well plates and 24 h later exposed to vehicle or P5091 at the indicated doses, in presence or absence of Z-VAD-FMK (20 μM), for 144 h and analysed for viability using a modifeid 3-(4,5-dimethylthiazole-2-yl)-2-5-diphenyltetrazolium bromide assay. CellTiter 96 Aqueous One Solution assay (Promega), as 50% inhibitory concentration (IC50) values. The value are presented as mean standard deviation of three independent experiments. Surviving fraction of LNCaP and PC3 cells are shown. (E) Caspase 3 activity was evaluated in LNCaP and PC3 cells treated or not treated with P5091 for 24 h, as indicated. The plotted values represent the mean +/− s.e.m. of three independent experiments.