Doxorubicin treatment modulates chemoresistance and affects the cell cycle in two canine mammary tumour cell lines

Autor: Barbara Brunetti, Cecilia Gola, Antonio Brocco, Monica Forni, Cinzia Benazzi, Asia Marangio, Chiara Bernardini, Raffaella De Maria, Augusta Zannoni, Roberta Salaroli, Giuseppe Sarli, Luisa Vera Muscatello, Federico Parenti, Michela Levi
Přispěvatelé: Levi M., Salaroli R., Parenti F., De Maria R., Zannoni A., Bernardini C., Gola C., Brocco A., Marangio A., Benazzi C., Muscatello L.V., Brunetti B., Forni M., Sarli G.
Rok vydání: 2020
Předmět:
p53
Proliferation index
Population
Canine mammary tumour
Mammary Neoplasms
Animal
P-glycoprotein
03 medical and health sciences
0302 clinical medicine
Dogs
Cell Line
Tumor
ATP Binding Cassette Transporter
Subfamily G
Member 2
Animals
Telomerase reverse transcriptase
ATP Binding Cassette Transporter
Subfamily B
Member 1
education
BCRP
Cell cycle
Cell line
Chemoresistance
Doxorubicin
Telomerases
030304 developmental biology
Cell Proliferation
0303 health sciences
education.field_of_study
lcsh:Veterinary medicine
General Veterinary
biology
Cell growth
Chemistry
Cell Cycle
General Medicine
Neoplasm Proteins
Gene Expression Regulation
Neoplastic
Apoptosis
Drug Resistance
Neoplasm
030220 oncology & carcinogenesis
Cancer research
biology.protein
Neoplastic cell
lcsh:SF600-1100
Multidrug Resistance-Associated Proteins
Research Article
Zdroj: BMC Veterinary Research
BMC Veterinary Research, Vol 17, Iss 1, Pp 1-15 (2021)
ISSN: 1746-6148
Popis: Background Doxorubicin (DOX) is widely used in both human and veterinary oncology although the onset of multidrug resistance (MDR) in neoplastic cells often leads to chemotherapy failure. Better understanding of the cellular mechanisms that circumvent chemotherapy efficacy is paramount. The aim of this study was to investigate the response of two canine mammary tumour cell lines, CIPp from a primary tumour and CIPm, from its lymph node metastasis, to exposure to EC50(20h) DOX at 12, 24 and 48 h of treatment. We assessed the uptake and subcellular distribution of DOX, the expression and function of P-glycoprotein (P-gp) and Breast Cancer Resistance Protein (BCRP), two important MDR mediators. To better understand this phenomenon the effects of DOX on the cell cycle and Ki67 cell proliferation index and the expression of p53 and telomerase reverse transcriptase (TERT) were also evaluated by immunocytochemistry (ICC). Results Both cell lines were able to uptake DOX within the nucleus at 3 h treatment while at 48 h DOX was absent from the intracellular compartment (assessed by fluorescence microscope) in all the surviving cells. CIPm, originated from the metastatic tumour, were more efficient in extruding P-gp substrates. By ICC and qRT-PCR an overall increase in both P-gp and BCRP were observed at 48 h of EC50(20h) DOX treatment in both cell lines and were associated with a striking increase in the percentage of p53 and TERT expressing cells by ICC. The cell proliferation fraction was decreased at 48 h in both cell lines and cell cycle analysis showed a DOX-induced arrest in the S phase for CIPp, while CIPm had an increase in cellular death without arrest. Both cells lines were therefore composed by a fraction of cells sensible to DOX that underwent apoptosis/necrosis. Conclusions DOX administration results in interlinked modifications in the cellular population including a substantial effect on the cell cycle, in particular arrest in the S phase for CIPp and the selection of a subpopulation of neoplastic cells bearing MDR phenotype characterized by P-gp and BCRP expression, TERT activation, p53 accumulation and decrease in the proliferating fraction. Important information is given for understanding the dynamic and mechanisms of the onset of drug resistance in a neoplastic cell population.
Databáze: OpenAIRE
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