Article Figures & Data
Figures
- Figure 1.
Metabolic features of isogenic 67NR and 4T1 breast cancer cells. A, LDH-A mRNA expression in 67NR and 4T1 cells by the semiquantitative RT-PCR. B, LDH-A, HXKII, and PKM2 proteins expression assessed by Western blotting. C, total LDH enzyme activity in 67NR cells and 4T1 cells. D, cell culture medium acidification; supernatant pH. E, oxygen consumption of 67NR and 4T1 cells in normal growth media [Dulbecco's Modified Eagle's Media (DMEM; high glucose) + 10% fetal calf serum + 2 mmol/L l-glutamine, 25 mmol/L HEPES] and upon treatment with 1 μmol/L of rotenone using the OxyLite system. F, glucose utilization; clearance of glucose from the culture medium. G, lactate production; appearance of lactate in the culture medium.
- Figure 2.
Lactate CSI spectra. A, different sizes of 67NR nonmetastatic tumors are shown. B, 4T1 metastasis-prone tumors are shown. Lactate signal was detected using CSI and a SelMQC editing sequence. C, LDH-A protein expression was assessed by Western blotting in different sizes of 67NR and 4T1 tumors. TV, tumor volume.
- Figure 3.
MRSI measurements of lactate concentration in orthotopic 67NR and 4T1 breast tumors during tumor growth. Lactate concentration in (A) 67NR tumors (solid circles, dashed line) and in (B) 4T1 tumors (solid squares, solid line) plotted versus tumor volume. C, average lactate concentration at 3 different tumor volumes (small, <150 mm3; medium, 150–400 mm3; and large, >400 mm3). Values are the mean ± SD. *, P < 0.05; **, P < 0.01. D, tumor lactate concentration plotted versus days postorthotopic implantation.
- Figure 4.
FDG-PET measurements of [18F]FDG accumulation in orthotopic 67NR and 4T1 breast tumors during tumor growth. [18F]FDG accumulation in (A) 67NR tumors (solid circles, dashed line) and in (B) 4T1 tumors (solid squares, solid line) plotted versus tumor volume. C, average [18F]FDG accumulation at 3 different tumor volumes (small, <150 mm3; medium, 150–400 mm3; and large, >400 mm3). Values are the mean ± SD. D, representative [18F]FDG microPET images of tumor-bearing mice; small and large 4T1 orthotopic tumors are visualized (white arrows). High radioactivity is seen in the cervical brown fat, heart, and bladder. Values are color coded to a range of values (%ID/mL).
- Figure 5.
The effect of hypoxia on the cell growth and LDH-A expression. A, growth profiles of 67NR cells and 4T1 cells under normoxic (N, 21%) and hypoxic (H, 1%) conditions. B, immunoblot analysis of LDH-A and LDH-B subunits from 67NR and 4T1 cells exposed to normoxia and hypoxia for 24 hours. C, the β-actin–normalized intensity of LDH-A and LDH-B levels in cells under normoxic and hypoxic conditions. D, the ratio of β-actin–normalized LDH-A and LDH-B expression under normoxic and hypoxic conditions.
- Figure 6.
Immunohistochemical staining of 67NR and 4T1 tumors. Histology (H&E), endothelial cell density (CD31, red), blood flow (Hoechst 33342, blue), and hypoxia (Pimonidazole, green) are compared. Small 67NR tumors showed no necrosis, a uniform endothelial cell density and blood flow, and no hypoxia. Medium-size 67NR tumors showed some pimonidazole staining. This small 4T1 tumor shows some central necrosis, a decrease in endothelial cell density, and blood flow, with a corresponding zone of hypoxia surrounding the necrotic zone. These observations were amplified in medium-size 4T1 tumors.
Additional Files
Supplementary Data
Files in this Data Supplement:
- Supplementary Figure 1 - PDF file - 442K
- Supplementary Figure 2 - PDF file - 192K
- Supplementary Figure 3 - PDF file - 790K
- Supplementary Figure 4 - PDF file - 1.05MB
- Supplementary Figure 5 - PDF file - 326K
Volume 17, Issue 19
