Fig. 1
Effects of masitinib treatment on cell viability of CMT-U309 and CMT-U27 cells. Data are expressed as mean percentage of cell viabilities ± standard error (SE) from three individual experiments. * -P < 0.05; ** -P < 0.01; *** -P < 0.001 compared to control
Table 1
IC20, IC50 and IC80 values of masitinib in CMT-U27 and CMT-U309 cells as measured by the MTT assay
| Cell line | Treatment | Time | IC20 (μM) | IC50 (μM) | IC80 (μM) |
|---|---|---|---|---|---|
| 24 h | 2.00 | 9.129 | 41.668 | ||
| CMT-U27 | Masitinib | 48 h | 3.099 | 7.607 | 18.670 |
| 72 h | 2.868 | 7.498 | 19.60 | ||
| 24 h | 2.497 | 15.032 | 90.485 | ||
| CMT-U309 | Masitinib | 48 h | 3.097 | 8.871 | 25.405 |
| 72 h | 2.471 | 8.545 | 29.544 |
Fig. 2
Effects of masitinib on DNA fragmentation of CMT-U27 and CMT-U309 cells. Data are expressed as mean OD values ± standard error from three individual experiments. * – P < 0.05; ** – P < 0.01; *** – P < 0.001 compared to control
Fig. 3
Flow cytometric analysis after incubation with masitinib (0–8 μM) for 72 h as representative profiles of annexin-V-FITC/PI staining of CMT-U27 cells and CMT-U309 cells. The lower left quadrant of the histogram shows viable cells (unstained by either fluorochrome) and the lower right one represents early apoptotic (annexin-V-positive) cells, indicating the translocation of phosphatidylserine to the external cell surface. The upper right quadrant represents late apoptotic (annexin-V- and PI-positive) cells, and the upper left one shows necrotic (PI-positive) cells. The numbers represent the mean percentage of cells (%) ± standard error
Fig. 4
Effects of masitinib treatment on the percentage of the total cell population in each phase of the cell cycle of CMT-U27 and CMT-U309 cells
Fig. 5
Immunocytochemical staining of Ki-67 in CMT-U27 and CMT-U309 cell lines. Bar = 20 μm. A – negative control, CMT-U27; B – negative control, CMT-U309; C – control high immunopositivity of Ki-67 in CMT-U27; D – control, high immunopositivity of Ki-67 in CMT-U309; E – IC50 masitinib, low immunopositivity of Ki-67 in CMT-U27; F – IC50 masitinib, low immunopositivity of Ki-67 in CMT-U309
Fig. 6
Effects of masitinib treatment on the proliferation index of CMT-U27 and CMT-U309 cells measured from Ki-67-stained slides
Table 2
VEGF concentrations in the supernatant of control and masitinib treated CMT cells after 72 h
| Masitinib concentration | CMT-U27 VEGF concentration (pg/mL) | CMT-U309 VEGF concentration (pg/mL) |
|---|---|---|
| Control | 536.5 ± 19.01 | 287.8 ± 14.25 |
| 0.25 μM | 504.64 ± 21.92 | 247.38 ± 14.98 |
| 0.5 μM | 439.48 ± 13.66* | 221.79 ± 10.32** |
| 1 μM | 440.28 ± 23.13* | 167.59 ± 8.05*** |
| 2 μM | 405.72 ± 21.05** | 154.90 ± 13.63*** |
| 4 μM | 399.87 ± 6.94*** | 99.59 ± 16.38*** |
| 8 μM | 102.74 ± 10.62*** | 67.97 ± 3.86*** |