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Effects of tyrosine kinase inhibitor-masitinib mesylate on canine mammary tumour cell lines

Fulya Ustun-Alkan
Fulya Ustun-Alkan
, 
Tülay Bakırel
Tülay Bakırel
, 
Oya Üstüner
Oya Üstüner
, 
Ceren Anlas
Ceren Anlas
, 
Suzan Cinar
Suzan Cinar
, 
Funda Yıldırım
Funda Yıldırım
 oraz 
Aydın Gürel
Aydın Gürel
   | 24 lip 2021
Journal of Veterinary Research's Cover Image
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Data publikacji: 24 lip 2021
Zakres stron: 351 - 359
Otrzymano: 05 lut 2021
Przyjęty: 05 lip 2021
DOI: https://doi.org/10.2478/jvetres-2021-042
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© 2021 F. Ustun-Alkan et al. published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

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
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

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
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
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
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
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
Effects of masitinib treatment on the proliferation index of CMT-U27 and CMT-U309 cells measured from Ki-67-stained slides

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***

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
eISSN:
2450-8608
Język:
Angielski
Częstotliwość wydawania:
4 razy w roku
Dziedziny czasopisma:
Life Sciences, Molecular Biology, Microbiology and Virology, other, Medicine, Veterinary Medicine
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