Figure 5

NDRG2 decreases the glucose uptake and GLUT1 protein levels in SK-BR-3-based subcutaneously xenograft tumours. The experiments illustrated are described in the Methods section. (A) Tumour growth was assessed every 3 days until day 21 treatment by measuring two perpendicular diameters and calculating the volume in cubic centimetres. Ad-LacZ, adenovirus expressing LacZ; Ad-NDRG2, adenovirus expressing NDRG2; PFU, Plaque-forming units. The data presented are means ± SD; error bars represent SD from 6 mice. *P < 0.05 and **P < 0.01 versus phosphate-buffered saline (PBS) or Ad-LacZ. (B) Tumour cells were dissociated from xenograft tumours and suspended in PBS after the number of cells was counted. Next, the glucose uptake of cells in each group was detected. The data presented are means ± SD of three independent experiments; error bars represent SD from 6 mice. *P < 0.05 and **P < 0.01 versus PBS or Ad-LacZ. (C) Intratumoural protein expression was assessed by N-myc downstream-regulated gene 2 (NDRG2) and glucose transporter 1 (GLUT1) IHC staining. Representative images are shown. Original magnification: 400 x; Scale bars = 50 μm. (D) Proteins of the xenograft tumours from each group were extracted and analysed by immunoblotting to quantify NDRG2 and GLUT1 protein changes.