Figure 1. VHL-deficiency augments the cytotoxicity of anthracyclines in ccRCC.

(a) RCC4/EV and RCC4/VHL were incubated in normoxia or hypoxia for 24 h and the indicated proteins expression was detected by western blot. (b) Different chemotherapeutic drugs were applied to treat RCC4/EV and RCC4/VHL cells for 24 h and cell growth inhibition rates were detected by CCK-8. (c) Doxorubicin was used to treat different ccRCC cell lines for 24 h and cell growth inhibition rates were detected and analysed using one-way ANOVA test followed by Bonferroni correction for post hoc test. ψ, # and , respectively, represent the difference of RCC4, 786-O and A498 versus OS-RC-2, Caki-2 , HK-2, Caki-1 or ACHN. (ψ, # and , P<0.05, ψψ and , P<0.01). (d,e) Western blot for VHL with actin as a loading control (bottom) and cell growth inhibition rates after treatment with doxorubicin or 1 μM VCR for 24 h (Top). (d) 786-O cells were stably transfected with VHL expression vector (VHL) or EV. (e) Caki-1 cells were infected with retroviral vectors harbouring shRNAs against VHL (shVHL) or NC. The column represents mean with bar as s.d. of three independent experiments with triplicate samples. (*P<0.05,**P<0.01 for t-test). ANOVA, analysis of variance; EPI, epirubicin; EV, empty vector; CDDP, cis-Diaminedichloroplatinum; CPT-11, irinotecan; DOX, doxorubicin and; DNR, daunomycin; 5-Fu, fluorouracil; NC, non-specific control; PS-341, bortezomib; TPT, topotecan; VCR, vincristine; VP-16, vepeside.