Figure 5

Effect of MCT1 silencing in vitro and in vivo. A) WI-38 conditioned medium recovered from fibroblasts treated with PC3-CM (WI38C CM) were added to PC3 and LNCaP cells cultured for 48 h in medium containing 0.6 g/L glucose. Proliferation assay was performed measuring the number of viable cells in control-silenced (CTR-sil) PCa cells, MCT1-silenced (MCT1-sil) PCa cells and using both WI38C CM and control WI-38 conditioned medium (WI38 CM). PCa cells cultured with WI38 CM were also treated with 5mM metformin. In blots time-course MCT1 expression in control and silenced PCa cells used in the proliferation assays was analyzed. Beta-tubulin detection was utilized as loading control. B) LNCaP and PC3 cells were injected alone or together with WI-38 cells s.c. in the flank of immunodeficient mice. Tumor growth was monitored daily and tumor growth was measured by a caliper. Experimental groups included: PCa cells alone; PCa cells injected with WI-38 fibroblasts; PCa cells injected with PC3-CM conditioned WI-38 fibroblasts (WI38C); MCT1-silenced PCa cells injected with WI38C; PCa cells injected with WI38C and treated with 50mg/kg metformin. *P>0.01 according to Student’s t test between PC3+WI38C and PC3+WI38 series. C) representative images of IHC analysis of xenograft tissue from mice inoculated with PC3 and WI38C (magnification 100x).