Article Figures & Data
Figures
- Figure 1.
GLPG1690 inhibited ATX activity and decreased LPA concentration in plasma. A, The structure of GLPG1690. B, Mice were treated for 5 days with a daily dose of 50 mg/kg or with 100 mg/kg GLPG1690 every 12 hours. The time point at 0 hour was obtained from mice that were treated with the vehicle for 5 days. Blood was collected by a terminal cardiac puncture at the times indicated from the first dose of GLPG1690 on day 5. For the 24-hour time point for the mice treated with 100 mg/kg GLPG1690, the second dose was given at 12 hours. C, Plasma concentrations of GLPG1690 from the mice treated with 100 mg/kg GLPG1690. The second curve, which is indicated, is the profile expected from repeating the dose at 12 hours. D–J, Plasma concentrations of different molecular species of LPA and of S1P and sphinganine 1-phosphate from the mice treated with 100 mg/kg GLPG690. n = 5 mice for the control and 3 mice in each treated group. *, P < 0.05; **, P < 0.01; ***, P < 0.001 compared with 0 hour for the lysophospholipids.
- Figure 2.
Effects of RT and GLPG1690 on breast tumor growth. A, Illustration of experiment using RT and GLPG1690 in mouse 4T1 breast tumor model. B, RT with or without GLPG1690 (GLPG) significantly decreased tumor growth. C, RT with or without GLPG1690 (GLPG) significantly decreased tumor weight at day 19 after injection of cancer cells. D, RT significantly decreased the percentage of Ki67-positive cells in tumors at day 19, which was decreased further by combination with GLPG1690. n = 5 mice for control, n = 6 mice other groups. *, P < 0.05 compared with control.
- Figure 3.
Effects of RT and GLPG1690 on [18F]FLT uptake by tumors. A and B, Representative static coronal [18F]FLT-PET images after 60 minutes postinjection during and after RT (7.5 Gy × 5 fractions). C and D, Quantitative [18F]FLT tumor uptake under the different experimental conditions as SUV from 5 or 6 mice in the control group and n = 6 mice in other groups. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
- Figure 4.
Effects of RT and GLPG1690 on mRNA levels of LPPs, LPA receptors, and ATX in tumors. A–C, mRNA levels of LPP1, LPP2, and LPP3. D–G, mRNA levels of LPA1, LPA2, LPA3, and LPA6. H, mRNA level of ATX. *, P < 0.05; **, P < 0.01; ***, P < 0.001 for 5 control mice and 6 mice in experimental groups.
- Figure 5.
Effects of RT and GLPG1690 on Bcl-2 and cleaved caspase-3 levels in tumors. A, RT with five daily fractions of 7.5 Gy or GLPG1690 treatment (100 mg/kg, every 12 hours) increased cleavage of caspase-3 and decreased Bcl-2 levels in tumors. B and C, Quantification of Western blotting for Bcl-2 and cleaved caspase-3. Samples from n = 5 control mice and n = 6 mice for other groups. *, P < 0.05.
- Figure 6.
Effects of RT and GLPG1690 on levels of inflammatory cytokines in tumors and tumor-adjacent adipose. Protein levels of CCL11 (A) in tumors and protein levels of IL9 (B), IL12 p40 (C), IFNγ (D), and M-CSF (E) in tumor-adjacent adipose tissue (TA) with or without five daily fractions of 7.5 Gy X rays and/or GLPG1690 (100 mg/kg, every 12 hours). Samples from n = 5 control mice and n = 6 mice for other groups. *, P < 0.05.
- Figure 7.
GLPG1690 increases the efficiency of doxorubicin in mouse model of breast cancer. A, Illustration of experiment using combination therapy with doxorubicin and GLPG1690 in mouse 4T1 breast tumor model. B and C, Doxorubicin (4 mg/kg, once every 2 days) in combination with GLPG1690 (100 mg/kg, every 12 hours) significantly decreased tumor growth and weight. D, Doxorubicin combined with GLPG1690 significantly decreased the percentage of Ki67-positive cells in tumors. E and F, Doxorubicin combined with GLPG1690 significantly increased 4-HNE-protein adducts in tumors. n = 6 mice from each group; *, P < 0.05 and **, P < 0.01 compared with control.
Additional Files
Supplementary Data
- Figure S1 - Body weight of mice A: Body weight of mice before and after treatment with GLPG1690 in combination with radiation. n = 5 to 6 mice per group. B: Body weight of mice before and after treatment with GLPG1690 in combination with doxorubicin. n = 6 mice per group.
- Figure S2 - Immunohistochemistry images for Ki67. A: Ki67 staining in tumors treated with GLPG1690 in combination with RT. B: Ki67 staining in tumors treated with GLPG1690 in combination with doxorubicin. Scale bar = 100 mm.
- Figure S3 - H&E and immunohistochemistry staining for ATX and Bcl-2 in mouse breast tumor tissue. The border between tumor (red T) and stromal tissue (red S) is showed as red dash line. Scale bar = 100 mm.
- Figure S4 - Immunohistochemistry staining for ATX and Bcl-2 in mouse breast tumor and tumor-adjacent adipose. A: ATX staining in tumors. B: ATX staining in tumor-adjacent adipose. C: Bcl-2 staining in tumors. Scale bar = 100 mm.
- Figure S5 - ATX activity in conditioned media of Hs578Bst stromal fibroblasts and patient-matching Hs578T breast cancer cells. Cells were exposed to different doses of g-radiation and ATX activity was measured 24 h after irradiation. n=3 to 5 for each condition. *P<0.05 compared with Hs578T cells.
- Figure S6 - Protein levels of cytokines in tumors with or without irradiation and/or GLPG1690. n = 5 in control, n = 6 in other groups. * P<0.05, ** P<0.01, ***P<0.001.
- Figure S7 - Protein levels of cytokines in tumor adjacent adipose tissue with or without irradiation and/or GLPG1690. n = 5 in control, n = 6 in other groups.
Volume 19, Issue 1
