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Relative quantification of DNA-Topoisomerases enzymes in cell lines: Correlation between cytotoxicity and mechanism of action of coordination compounds

Abstract

DNA-topoisomerases enzymes are one of main biological targets against cancer. These proteins participate of different nuclear processes. Literature shows that some types of cancer have a great expression of the topoisomesae I² (TOPO1B) and/or topoisomesae II± (TOPO2A). However, few studies were performed to prove the quantities of the enzymes inside the cells. Thus, this Project has the objective to determine the relative quantities of the topoisoemrases enzymes by Western Blot e ELISA methodologies in different cells lines: MDA-MB-231 (metastatic breast cancer), MCF7 (breast cancer), DU 145 (prostate cancer), A549 (lung cancer), A2780cis (cisplatin-resistant ovary cancer), MCF10A (non-tumoral breast), PNT2 (non-tumoral prostate), MCR5 (non-tumoral lung). Additionally, we intend to synthesize, characterize and evaluate the cytotoxicity and the capacity to inhibit the action of TOPO1B and TOPO2A of a large number of coordination compounds with different metal ions (Pd2+, Pt2+ e Ag+). A deep investigation of mechanism of action of the most promising compounds will be making by cell cycle arrest and apoptosis assays. To determine the concentration of metal íon inside the cellular media ICP-MS analysis would be performed. Therefore, we expect that these results help us to explain experimentally the relationship between the expressions of the enzymes, cytotoxicity and mechanism of action. Indeed, our works will achieve another scientific level. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
HOOPER, CATHERINE A. J.; CARDO, LUCIA; CRAIG, JAMES S.; MELIDIS, LAZAROS; GARAI, ADITYA; EGAN, ROSS T.; SADOVNIKOVA, VIKTORIIA; BURKERT, FLORIAN; MALE, LOUISE; HODGES, NIKOLAS J.; BROWNING, DOUGLAS F.; ROSAS, ROSELYNE; LIU, FENGBO; ROCHA, V, FILLIPE; LIMA, MAURO A.; LIU, SIMIN; BARDELANG, DAVID; HANNON, MICHAEL J. Rotaxanating Metallo-supramolecular Nano-cylinder Helicates to Switch DNA Junction Binding. Journal of the American Chemical Society, v. 142, n. 49, p. 20651-20660, DEC 9 2020. Web of Science Citations: 0.
DE MOURA, THALES REGGIANI; ZANETTI, RENAN DIEGO; SILVA, DEBORA EDUARDA SOARES; DE FARIAS, RENAN LIRA; MAURO, ANTONIO EDUARDO; PEREIRA, JOSE CLAYSTON MELO; DE SOUZA, ALINE APARECIDA; DA SILVA SIQUEIRA, FABIO; DE SOUZA JUDICE, WAGNER ALVES; LIMA, MAURO ALMEIDA; ROCHA, FILLIPE VIEIRA; DEFLON, VICTOR MARCELO; VIEIRA DE GODOY NETTO, ADELINO. Palladium(ii) complexes bearing 1-iminothiolate-3,5-dimethylpyrazoles: synthesis, cytotoxicity, DNA binding and enzymatic inhibition studies. NEW JOURNAL OF CHEMISTRY, v. 44, n. 45, p. 19891-19901, DEC 7 2020. Web of Science Citations: 0.
LIMA, MAURO ALMEIDA; COSTA, VINICIUS A.; FRANCO, MARIANE ARALIJO; DE OLIVEIRA, GABRIELA P.; DEFLON, VICTOR M.; ROCHA, FILLIPE VIEIRA. Palladium(II) complexes bearing thiosemicarbazone and phosphines as inhibitors of DNA-Topoisomerase II enzyme: Synthesis, characterizations and biological studies. Inorganic Chemistry Communications, v. 112, FEB 2020. Web of Science Citations: 1.

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