Proteomics Approach of Rapamycin Anti-Tumoral Effect on Primary and Metastatic Canine Mammary Tumor Cells In Vitro

Autor(es)

Lainetti, Patricia F. ^[1] ; Leis-Filho, Antonio F. ^[2] ; Kobayashi, Priscila E. ^[2] ; de Camargo, Laiza S. ^[1] ; Laufer-Amorim, Renee ^[2] ; Fonseca-Alves, Carlos E. ^{[1, 3]} ; Souza, Fabiana F. ^[1]

Afiliação do(s) autor(es)

^[1] Sao Paulo State Univ UNESP, Sch Vet Med & Anim Sci, Dept Vet Surg & Anim Reprod, BR-18618681 Botucatu, SP - Brazil

^[2] Sao Paulo State Univ Unesp, Sch Vet Med & Anim Sci, Dept Vet Clin, BR-18618681 Botucatu, SP - Brazil

^[3] Univ Paulista UNIP, Inst Hlth Sci, BR-17048290 Bauru, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Molecules; v. 26, n. 5 MAR 2021.
• Citações Web of Science: 0

Resumo

Rapamycin is an antifungal drug with antitumor activity and acts inhibiting the mTOR complex. Due to drug antitumor potential, the aim of this study was to evaluate its effect on a preclinical model of primary mammary gland tumors and their metastases from female dogs. Four cell lines from our cell bank, two from primary canine mammary tumors (UNESP-CM1, UNESP-CM60) and two metastases (UNESP-MM1, and UNESP-MM4) were cultured in vitro and investigated for rapamycin IC50. Then, cell lines were treated with rapamycin IC50 dose and mRNA and protein were extracted in treated and non-treated cells to perform AKT, mTOR, PTEN and 4EBP1 gene expression and global proteomics by mass spectrometry. MTT assay demonstrated rapamycin IC50 dose for all different tumor cells between 2 and 10 mu M. RT-qPCR from cultured cells, control versus treated group and primary tumor cells versus metastatic tumor cells, did not shown statistical differences. In proteomics were found 273 proteins in all groups, and after data normalization 49 and 92 proteins were used for statistical analysis for comparisons between control versus rapamycin treatment groups, and metastasis versus primary tumor versus metastasis rapamycin versus primary tumor rapamycin, respectively. Considering the two statistical analysis, four proteins, phosphoglycerate mutase, malate dehydrogenase, l-lactate dehydrogenase and nucleolin were found in decreased abundance in the rapamycin group and they are related with cellular metabolic processes and enhanced tumor malignant behavior. Two proteins, dihydrolipoamide dehydrogenase and superoxide dismutase, also related with metabolic processes, were found in higher abundance in rapamycin group and are associated with apoptosis. The results suggested that rapamycin was able to inhibit cell growth of mammary gland tumor and metastatic tumors cells in vitro, however, concentrations needed to reach the IC50 were higher when compared to other studies. (AU)

• Processo FAPESP: 19/24079-1 - Efeito antitumoral do látex Synadenium grantii Hook F. em modelos pré-clínicos de carcinomas mamários
• Beneficiário: Carlos Eduardo Fonseca Alves
• Modalidade de apoio: Auxílio à Pesquisa - Regular