Figure 4
HIF-1-mediated metabolic reprogramming reduced ROS levels and increased the survival of metastatic cancers.
Athymic nude mice were i.v. transplanted with a suspension of the stable transfectant, EMT6/CMVp-CLuc/5HREp-FLuc cells (1 × 106/mouse) and treated with or without the HIF-1 inhibitor, YC-1 (30 mg/kg daily) from 3 to 6 days after the transplantation of cancer cells. (a) Lungs with the resultant metastatic tumors were surgically excised 7 days after the transplantation, which was 1 day after the peak in HIF-1 activity and ROS levels (SeeFig. 1h,2b) and ROS was detected in methacarn-fixed paraffin-embedded sections (upper) and formalin-fixed paraffin-embedded sections (lower) by the Protein Carbonyls Immunohistochemical Staining Kit and anti-phospho-p38 antibody, respectively. Bar = 500 μm (upper) and 20 μm (lower). (b) Ten days after the transplantation, mice were subjected to the in vivo CLuc assay in the serum for tumor volume. (c) Lungs with the resultant metastatic tumors were surgically excised 10 days after the transplantation, treated with Bouin's solution and the number of colonies was externally counted. Means ± s.d. n = 10. *P < 0.05. (d) Representative images of the lungs are shown.
