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Fig. 3 | Journal of Experimental & Clinical Cancer Research

Fig. 3

From: p53 mutant-type in human prostate cancer cells determines the sensitivity to phenethyl isothiocyanate induced growth inhibition

Fig. 3

PEITC inhibits cell proliferation, induces apoptosis and reactivates p53P223L/V274F mutant in DU145 cells. Prostate cancer cells with p53 mutations p53P223L/V274F (DU145), WT p53 (LNCaP), p53 null (PC-3) and normal prostate epithelial cells (RWPE2) were treated with DMSO or PEITC for 24 h. a Percent cell proliferation was determined by the WST-1 assay, and b Apoptosis was measured by Annexin-V staining by flow cytometry using a BD LSRFORTESSA instrument. (*p ≤ 0.02). c Immunoprecipitation of the p53 mutant protein from DU145 cell lysates by using p53 mutant-specific antibody PAB240 and detected by a general anti-p53 (FL393) antibody. The PC-3 (p53 null) cells were used as a control for PAB240 antibody. Input lysates were probed with a general anti-p53 (DO-1) antibody. Blots were stripped and reprobed with anti-GAPDH antibody. d DU145 cells were treated with PEITC for 4 h and chromatin-bound and soluble nuclear fractions were analyzed by immunoblotting. Histone H3 and Topoisomerase IIB served as markers for the chromatin and soluble nuclear fractions, respectively. e qRT-PCR of p53 regulated genes in DU145, LNCaP and PC-3 cells treated with DMSO or 8 μM PEITC for 4 h. (***p ≤ 0.0009 and **p ≤ 0.006). f DU145 cells were treated with DMSO or the indicated concentration of PEITC for 4 h. The cell lysate was resolved by SDS-PAGE, probed with p53 DO-1 antibody and re-probed with anti-GAPDH antibody

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