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

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

Journal of Veterinary Research
Volume 65 (2021): Issue 3 (September 2021)
By: Fulya Ustun-Alkan,  Tülay Bakırel,  Oya Üstüner,  Ceren Anlas,  Suzan Cinar,  Funda Yıldırım and  Aydın Gürel  
Open Access
|Jul 2021

Figures & Tables

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 concentrationCMT-U27 VEGF concentration (pg/mL)CMT-U309 VEGF concentration (pg/mL)
Control536.5 ± 19.01287.8 ± 14.25
0.25 μM504.64 ± 21.92247.38 ± 14.98
0.5 μM439.48 ± 13.66*221.79 ± 10.32**
1 μM440.28 ± 23.13*167.59 ± 8.05***
2 μM405.72 ± 21.05**154.90 ± 13.63***
4 μM399.87 ± 6.94***99.59 ± 16.38***
8 μM102.74 ± 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 lineTreatmentTimeIC20 (μM)IC50 (μM)IC80 (μM)
24 h2.009.12941.668
CMT-U27Masitinib48 h3.0997.60718.670
72 h2.8687.49819.60
24 h2.49715.03290.485
CMT-U309Masitinib48 h3.0978.87125.405
72 h2.4718.54529.544
DOI: https://doi.org/10.2478/jvetres-2021-042 | Journal eISSN: 2450-8608
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Language: English
Page range: 351 - 359
Submitted on: Feb 5, 2021
|
Accepted on: Jul 5, 2021
|
Published on: Jul 24, 2021
Published by: National Veterinary Research Institute in Pulawy
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year
Keywords:
tyrosine kinase inhibitor,
masitinib,
canine mammary cancer,
apoptosis,
VEGF
Related subjects:
Life sciences,
Molecular biology,
Microbiology and virology,
Life sciences, other,
Medicine,
Veterinary medicine

© 2021 Fulya Ustun-Alkan, Tülay Bakırel, Oya Üstüner, Ceren Anlas, Suzan Cinar, Funda Yıldırım, Aydın Gürel, published by National Veterinary Research Institute in Pulawy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

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