Carboplatin effect on canine benign mixed- tumour-derived cells cultured under three-dimensional system: apoptosis, cell viability and mitochondrial dysfunction

Authors

DOI:

https://doi.org/10.31533/pubvet.v17n04e1368

Keywords:

apoptosis, carboplatin, celular culture 3D, MC-TMB

Abstract

Canine mammary carcinomas represent an important pathology in small animal clinics, and benign mixed type carcinomas (MC-BMT) are one of the most diagnosed worldwide. The use of chemotherapeutic carboplatin has been one of the new protocols for the treatment of BMT. In this respect, three-dimensional cell culture (3D) represents an alternative in the evaluation of drugs by simulating what occurs in vivo. The present study aimed to verify the effect of carboplatin on a 3D culture of cells derived from BMT, in addition to possible changes in cell viability, spheroid size, apoptosis and Bcl-2/Bax ratio. For this, tumour samples were collected during mastectomy procedures in private veterinary clinics, which were submitted to in vitro culture, and part of the collected fragments were submitted to histopathological analysis. After 28 days of 3D culture, spheroids were documented in both groups (treated and control) and sizes and morphology were compared. The carboplatin interfered in the cell viability by affecting their division and promoting apoptotic events. In the treated group, a higher transcription of Bax and caspase 3 was observed, in addition to low levels of caspases 2, 8 and 9, which was not observed in the control group. We thus suggest that mitochondrial dysfunction plays a critical role in cancer progression and that targeting mitochondrial alterations and mitochondrial retrograde signalling might be a promising strategy for the development of selective anticancer therapy. Thus, it was possible to demonstrate that the results achieved may contribute to the establishment of a new chemotherapy for female dogs with MC-BMT.

References

Alfarouk, K. O., Stock, C. M., Taylor, S., Walsh, M., Muddathir, A. K., Verduzco, D., Bashir, H. H., Mohammed, O. Y., Elhassan, G. O., Harguindey, S., Reshkin, S. J., Ibrahin, M. E. & Rauch, C. (2015). Resistance to cancer chemotherapy: failure in drug response from ADME to P-gp. Cancer Cell International, 15:1-13.‏https://doi.org/10.1186/s12935-015-0221-1.

Bertolini, F., Sukhatme, V. P. & Bouche, G. (2015). Drug repurposing in oncology - patient and health systems opportunities. Nature Reviews Clinical Oncology, 12:732-742.‏ https://doi.org/10.1038/nrclinonc.2015.169.

Cardoso, T. C., Sakamoto, S. S., Stockmann, D., Souza, T. F. B., Ferreira, H. L., Gameiro, R., Vieira, F. V., Louzada, M. J. Q., Andrade, A. L. & Flores, E. F. (2017). A three-dimensional cell culture system as an in vitro canine mammary carcinoma model for the expression of connective tissue modulators. Veterinary and Comparative Oncology, 15,582-593. https://doi.org/10.1111/vco.12202.

Cassali, G. D., Lavalle, G. E., Ferreira, E., Estrela-Lima, A., De Nardi, A. B., Ghever, C. et al. (2013). Consensus for the Diagnosis, Prognosis and Treatment of Canine Mammary Tumors. Brazilian Journal of Veterinary Pathology, 7:38-69. https://doi.org/10.24070/bjvp.1983-0246.v13i3p555-574.

Chee, J. L., Saidin, S., Lane, D. P., Leong, S. M., Noll, J. E., Neilsen, P. M., Phua, Y. T., Gabra, H. & Lim, T. M. (2013). Wild-type and mutant p53 mediate cisplatin resistance through interaction and inhibition of active caspase-9. Cell Cycle, 12:278-288. https://doi.org/10.4161/cc.23054.

Chia-Chi, H., Ling-Ming, T. & Hsin-Chen, L. (2016). Role of mitochondrial dysfunction in cancer progression. Experimental Biology and Medicine, 241:1281-1295. https://doi.org/10.1177/1535370216641787

Damasceno, K. A., Bertagnolli, A. C., Estrela-Lima, A., Ribeiro, L. G., Rabelo, B. S., Campos, C. B., Barros, A. L. & Cassali, G. D. (2012). Versican expression in canine carcinomas in benign mixed tumours: is there an association with clinical pathological factors, invasion and overall survival? BMC Veterinary Research, 8:195. https://doi.org/10.1186/1746-6148-8-195.

Estrela-Lima, A., Araújo, M. S., Costa-Neto, J. M., Teixeira-Carvalho, A., Barrouin-Melo, S. M., Cardoso, S. V., Martins-Filho, O. A., Serakides, R. & Cassali, G. D. (2010). Immunophenotypic features of tumor infiltrating lymphocytes from mammary carcinomas in female dogs associated with prognostic factors and survival rates. BMC Cancer, 10:256. https://doi.org/10.1186/1471-2407-10-256

Ferreira, E., Bregunci, G. C., Schmitt, F. C. & Cassali, G. D. (2003). Protocol for the anatomopathological examination of canine mammary tumors. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 55:105-109. https://doi.org/10.1590/s0102-09352003000100017.

Fisusi, F. A. & Akala, E. O. (2019). Drug combinations in breast cancer therapy. Pharmaceutical Nanotechnology, 7:3-23.‏ https://doi.org/10.2174/2211738507666190122111224.

Frengki, D. P. P., Wahyuni, F. S., Khambri, D & Sofia, V. (2021). The effect of deoxyelephantopin enhances doxorubicin sensitivity to mcf-7 cancer cells. Research Journal of Pharmacy and Technology, 14:2791-2795. https://doi.org/10.52711/0974-360x.2021.00492.

Jin, K. T., Lu, Z. B., Chen, J. Y., Liu, Y. Y., Lan, H. R., Dong, H. Y., Yang, F., Zhao, Y. Y. & Chen, X. Y. (2020). Recent trends in nanocarrier-based targeted chemotherapy: selective delivery of anticancer drugs for effective lung, colon, cervical, and breast cancer treatment. Journal of Nanomaterials. https://doi.org/10.1155/2020/9184284.

Lavalle, G. E., De Campos, C. B., Bertagnolli, A. C. & Cassali, G. D. (2012). Canine malignant mammary gland neoplasms with advanced clinical staging treated with carboplatin and cyclooxygenase inhibitors. In Vivo, 26:375-379.

Livak, K. J. & Schmittgen, T. D. (2001). Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2-∆∆CT Method. Methods, 25:402-408. https://doi.org/10.1006/meth.2001.1262.

Misdorp, W. (2002). Tumors of the mammary gland. Tumors in Domestic Animals (4th ed.). Iowa State: Blackwell Publishing.

Narayan, R. S., Molenaar, P., Teng, J., Cornelissen, F. M. G., Roelofs, I., Menezes, R., Dik, R., Lagerweij, T., Broersma, Y., Petersen, N., Marin Soto, J. A., Brands, E., van Kuiken, P., Lecca, M. C., Lenos, K. J., In 't Veld, S. G. J. G., van Wieringen, W., Lang, F. F., Sulman, E., Verhaak, R. & Westerman, B. A. (2020). A cancer drug atlas enables synergistic targeting of independent drug vulnerabilities. Nature Communications, 11:2935. https://doi.org/10.1038/s41467-020-16735-2.

Nowak-Sliwinska, P., Scapozza, L. & Altaba, A. R. (2019). Drug repurposing in oncology: Compounds, pathways, phenotypes and computational approaches for colorectal cancer. Biochimica et Biophysica Acta (BBA)-Reviews on Cancer, 1871:434-454.‏ https://doi.org/10.1016/j.bbcan.2019.04.005.

Owen, L. N. (1980). TNM Classification of Tumours in Domestic Animals/ edited by L. N. Owen. World Health Organization.

Rangel, M. M. M., Brito, C. P., Oliveira, K. D. & Dagli, M. L. Z. (2009). Fibrosarcoma recidivante em cão e eletroquimioterapia adjuvante como terapia de controle. Resumos, FMVZ/USP.

Salas, Y., MaÂrquez, A., Diaz, D. & Romero, L. (2015). Epidemiological study of mammary tumors in female dogs diagnosed during the period 2002-2012: A growing animal health problem. PLOS One, 10:e0127381. https://doi.org/10.1371/journal.pone.0127381.

Sousa, G. F. D., Wlodarczyk, S. R. & Monteiro, G. (2014). Carboplatin: molecular mechanisms of action associated with chemoresistance. Brazilian Journal of Pharmaceutical Sciences, 50:693-701.‏ https://doi.org/10.1590/s1984-82502014000400004.

Ulukaya, E., Colakogullari, M. & Wood, E. J. (2004). Interference by anti-cancer chemotherapeutic agents in the MTT-tumor chemosensitivity assay. Chemotherapy, 50:43-50.‏ https://doi.org/10.1159/000077285.

Uwano, M., Kano, R., Maruyama, H., Hasegawa, A. & Kamata, H. (2012). Therapeutic Efficacy of ABT-737, a Bcl-2 Inhibitor, in a Canine Melanoma Cell Line Journal Veterinary Medicine, 74:783-785. https://doi.org/10.1292/jvms.11-0431.

Vasan, N., Baselga, J. & Hyman, D.M. (2019). A view on drug resistance in cancer. Nature, 575,299-309.‏ https://doi.org/10.1038/s41586-019-1730-1.

Wang, X., Zhang, H., Chen, X. 2020. Drug resistance and combating drug resistance in cancer. Cancer Drug Resistance, 2:141-160. https://doi.org/10.20517/cdr.2019.10.

Published

2023-04-07

Issue

Section

Biotecnologia

How to Cite

Carboplatin effect on canine benign mixed- tumour-derived cells cultured under three-dimensional system: apoptosis, cell viability and mitochondrial dysfunction. (2023). Pubvet, 17(04), e1368. https://doi.org/10.31533/pubvet.v17n04e1368

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