Mecanismo de morte celular induzida por complexos de rutênio II e III em diferentes linhagens tumorais
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFG |
Texto Completo: | http://repositorio.bc.ufg.br/tede/handle/tede/4787 |
Resumo: | The resistance acquired by some tumor cell lines retricts the use of drugs made of platinum because of Ruthenium compounds have been objects of great attention for presenting antimetastatic properties and low toxicity. Ruthenium compounds form compounds with the most different chemical binders, presenting good behavior and expanding the possibilities offor biological applications. A wide variety of coordination has enabled studies on ruthenium complexes, andseveral oxidation stages (Ru (II), Ru (III), and Ru (IV)) under physiological conditions and the rate of binder substitution. This study ranges the citotoxic activity of ruthenium (III) compound cis- Tetraammine(oxalato)Ruthenium(III) Dithionate - {Cis-[Ru(C2O4)(NH3)4]2(S2O6)} to treat human erythroleukemia (K562) tumor cell lineand the complex of ruthenium (II) coordinated a phosphine ligand and nitrile front tumor lineage S180 through the techniques of assay cell viability, assay kinetics of cell cycle phases, annexin V assay/ propidium iodide, test of mitochondrial membrane potential, test comet and gene expression through real time PCR. Both antiproliferative and cytotoxic activity revealed that K562 cells cultured with ruthenium (III) compound showed meaningful decrease in proliferation. Ruthenium(III) compound induced the IC50 value was of 18.28 μM set againts the cell cycle profiles cells not treated. Flow cytometric analysis indicated a sub-G1 arresting effect of ruthenium compound on K562 cells. Through the cell viability assay through MTT reduction technique, it was found that the complex of ruthenium (II) phosphine coordinated and nitrile presented cytotoxic activity when facing the tumor strain S180 with IC50 17.02±8.21μM and IC50 de 53.73 ± 5.71 μM for lymphocyte. When analyzing the cell cycle of tumor cells S180 treated with complex of ruthenium (II) caused increase in cells in G0/G1 and in S phase decreased. We observed an increase G2 / M. In the analysis of apoptosis assays, the results pointed that the complex ruthenium (II) induced cell death via apoptosis in tumor strain S180 as proved the increase in annexin cells V positive, depolarization of the mitochondrial membrane potential, activation of caspase 3 (Casp3) and 8 (Casp8) and increased expression levels of caspase-3 (Casp3) (mRNA), Bax (mRNA) and Tp53. The results lead to the conclusion that both complexes of ruthenium (II) and (III) induce cytotoxic activity against cell models tested, and that this activity correlates with alterations in cell cycle phases and induction of cell death via apoptosis. |
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Lacerda, Elisangela de Paula Silveirahttp://lattes.cnpq.br/9390789693192751Lacerda, Elisangela de Paula SilveiraGouvea, Wagner dos SantosMenck, Carlos Frederico MartinsLobato, YandraSilva, Claudio Carlos dahttp://lattes.cnpq.br/2888523360892312Pereira, Flavia de Castro2015-10-27T14:38:45Z2014-03-19PEREIRA, F. C. Mecanismo de morte celular induzida por complexos de rutênio II e III em diferentes linhagens tumorais. 2014. 162 f. Tese (Doutorado em Biologia) - Universidade Federal de Goiás, Goiânia, 2014.http://repositorio.bc.ufg.br/tede/handle/tede/4787ark:/38995/0013000006d5gThe resistance acquired by some tumor cell lines retricts the use of drugs made of platinum because of Ruthenium compounds have been objects of great attention for presenting antimetastatic properties and low toxicity. Ruthenium compounds form compounds with the most different chemical binders, presenting good behavior and expanding the possibilities offor biological applications. A wide variety of coordination has enabled studies on ruthenium complexes, andseveral oxidation stages (Ru (II), Ru (III), and Ru (IV)) under physiological conditions and the rate of binder substitution. This study ranges the citotoxic activity of ruthenium (III) compound cis- Tetraammine(oxalato)Ruthenium(III) Dithionate - {Cis-[Ru(C2O4)(NH3)4]2(S2O6)} to treat human erythroleukemia (K562) tumor cell lineand the complex of ruthenium (II) coordinated a phosphine ligand and nitrile front tumor lineage S180 through the techniques of assay cell viability, assay kinetics of cell cycle phases, annexin V assay/ propidium iodide, test of mitochondrial membrane potential, test comet and gene expression through real time PCR. Both antiproliferative and cytotoxic activity revealed that K562 cells cultured with ruthenium (III) compound showed meaningful decrease in proliferation. Ruthenium(III) compound induced the IC50 value was of 18.28 μM set againts the cell cycle profiles cells not treated. Flow cytometric analysis indicated a sub-G1 arresting effect of ruthenium compound on K562 cells. Through the cell viability assay through MTT reduction technique, it was found that the complex of ruthenium (II) phosphine coordinated and nitrile presented cytotoxic activity when facing the tumor strain S180 with IC50 17.02±8.21μM and IC50 de 53.73 ± 5.71 μM for lymphocyte. When analyzing the cell cycle of tumor cells S180 treated with complex of ruthenium (II) caused increase in cells in G0/G1 and in S phase decreased. We observed an increase G2 / M. In the analysis of apoptosis assays, the results pointed that the complex ruthenium (II) induced cell death via apoptosis in tumor strain S180 as proved the increase in annexin cells V positive, depolarization of the mitochondrial membrane potential, activation of caspase 3 (Casp3) and 8 (Casp8) and increased expression levels of caspase-3 (Casp3) (mRNA), Bax (mRNA) and Tp53. The results lead to the conclusion that both complexes of ruthenium (II) and (III) induce cytotoxic activity against cell models tested, and that this activity correlates with alterations in cell cycle phases and induction of cell death via apoptosis.Fármacos à base de cisplatina ainda são os anticancerígenos mais utilizados no mundo. Os compostos de rutênio têm sido objetos de grande atenção devido as suas propriedades antimetastática, baixa toxicidade e vários estados de oxidação (Ru (II), Ru (III) e Ru (IIV)) em condições fisiológicas. O presente trabalho teve como objetivo investigar in vitro os efeitos citotóxico e mecanismo de morte do complexo Ditionato de cis-Tetraamino(oxalato)rutênio(III) na linhagem tumoral de leucemia mielóide crônica (K562) e do complexo de rutênio (II) coordenado a ligante fosfina e nitrila [RuCl(bcn)(bipy)(dppe)]PF6, em linhagem tumoral S180 a partir das técnicas de ensaio de viabilidade celular, ensaio de cinética das fases do ciclo celular, ensaio anexina V/Iodeto de Propídeo, ensaio de potencial de membrana mitocondrial e expressão gênica por PCR em tempo real. Os resultados mostraram que o complexo de rutênio (III) provoca uma significativa redução na proliferação de células K562. O complexo de rutênio (III) induziu uma IC50 =18,28 μM. A análise por citometria de fluxo indicou um efeito sub-G1 dos complexos de rutênio sobre células de K562. O composto provocou um aumento de dano significativo nas células em todas as concentrações testadas em comparação ao controle negativo, o que pode ser associado à citotoxicidade com efeito direto sobre o DNA das células de K562. A partir do ensaio de viabilidade celular pela técnica de redução do MTT, verificou-se que o complexo de rutênio (II) coordenado a fosfina e nitrila apresentou atividade citotóxica frente à linhagem tumoral S180 com IC50 17,02±8,21μM e IC50 de 53,73 ± 5,71 para linfócito. Na análise do ciclo celular de células tumorais S180 tratadas com o complexo de rutênio (II), casou indução de G0/G1, fase S e G2/M. Na análise dos ensaios de apoptose, os resultados demonstraram que o complexo de rutênio (II) induziu morte celular via apoptose na linhagem tumoral S180, como evidenciado pelo aumento no número de células anexina V positivo, despolarização do potencial de membrana mitocondrial, ativação das caspase 3 (Casp3) e 8 (Casp8) e aumento dos níveis de expressão de caspase-3 (Casp3) (mRNA), Bax (mRNA) e Tp53 . A partir dos resultados conclui-se que ambos os complexos de rutênio (II) e (III) induzem atividade citotóxica frente aos modelos de células testadas, sendo que a atividade está correlacionada às alterações nas fases do ciclo celular e indução de morte celular via apoptose.Submitted by Cláudia Bueno (claudiamoura18@gmail.com) on 2015-10-26T16:54:27Z No. of bitstreams: 2 Tese - Flávia de Castro Pereira - 2014.pdf: 7274098 bytes, checksum: 2478fc770e9bfbd8ffb6361d038b525d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2015-10-27T14:38:45Z (GMT) No. of bitstreams: 2 Tese - Flávia de Castro Pereira - 2014.pdf: 7274098 bytes, checksum: 2478fc770e9bfbd8ffb6361d038b525d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5)Made available in DSpace on 2015-10-27T14:38:45Z (GMT). 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dc.title.por.fl_str_mv |
Mecanismo de morte celular induzida por complexos de rutênio II e III em diferentes linhagens tumorais |
title |
Mecanismo de morte celular induzida por complexos de rutênio II e III em diferentes linhagens tumorais |
spellingShingle |
Mecanismo de morte celular induzida por complexos de rutênio II e III em diferentes linhagens tumorais Pereira, Flavia de Castro Apoptose Ciclo celular Citotoxicidade Complexos de rutenio II e III Apoptosis Cell cycle Cytotoxicity Ruthenium complexes II and III BIOQUIMICA::BIOLOGIA MOLECULAR MORFOLOGIA::CITOLOGIA E BIOLOGIA CELULAR |
title_short |
Mecanismo de morte celular induzida por complexos de rutênio II e III em diferentes linhagens tumorais |
title_full |
Mecanismo de morte celular induzida por complexos de rutênio II e III em diferentes linhagens tumorais |
title_fullStr |
Mecanismo de morte celular induzida por complexos de rutênio II e III em diferentes linhagens tumorais |
title_full_unstemmed |
Mecanismo de morte celular induzida por complexos de rutênio II e III em diferentes linhagens tumorais |
title_sort |
Mecanismo de morte celular induzida por complexos de rutênio II e III em diferentes linhagens tumorais |
author |
Pereira, Flavia de Castro |
author_facet |
Pereira, Flavia de Castro |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Lacerda, Elisangela de Paula Silveira |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/9390789693192751 |
dc.contributor.referee1.fl_str_mv |
Lacerda, Elisangela de Paula Silveira |
dc.contributor.referee2.fl_str_mv |
Gouvea, Wagner dos Santos |
dc.contributor.referee3.fl_str_mv |
Menck, Carlos Frederico Martins |
dc.contributor.referee4.fl_str_mv |
Lobato, Yandra |
dc.contributor.referee5.fl_str_mv |
Silva, Claudio Carlos da |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/2888523360892312 |
dc.contributor.author.fl_str_mv |
Pereira, Flavia de Castro |
contributor_str_mv |
Lacerda, Elisangela de Paula Silveira Lacerda, Elisangela de Paula Silveira Gouvea, Wagner dos Santos Menck, Carlos Frederico Martins Lobato, Yandra Silva, Claudio Carlos da |
dc.subject.por.fl_str_mv |
Apoptose Ciclo celular Citotoxicidade Complexos de rutenio II e III |
topic |
Apoptose Ciclo celular Citotoxicidade Complexos de rutenio II e III Apoptosis Cell cycle Cytotoxicity Ruthenium complexes II and III BIOQUIMICA::BIOLOGIA MOLECULAR MORFOLOGIA::CITOLOGIA E BIOLOGIA CELULAR |
dc.subject.eng.fl_str_mv |
Apoptosis Cell cycle Cytotoxicity Ruthenium complexes II and III |
dc.subject.cnpq.fl_str_mv |
BIOQUIMICA::BIOLOGIA MOLECULAR MORFOLOGIA::CITOLOGIA E BIOLOGIA CELULAR |
description |
The resistance acquired by some tumor cell lines retricts the use of drugs made of platinum because of Ruthenium compounds have been objects of great attention for presenting antimetastatic properties and low toxicity. Ruthenium compounds form compounds with the most different chemical binders, presenting good behavior and expanding the possibilities offor biological applications. A wide variety of coordination has enabled studies on ruthenium complexes, andseveral oxidation stages (Ru (II), Ru (III), and Ru (IV)) under physiological conditions and the rate of binder substitution. This study ranges the citotoxic activity of ruthenium (III) compound cis- Tetraammine(oxalato)Ruthenium(III) Dithionate - {Cis-[Ru(C2O4)(NH3)4]2(S2O6)} to treat human erythroleukemia (K562) tumor cell lineand the complex of ruthenium (II) coordinated a phosphine ligand and nitrile front tumor lineage S180 through the techniques of assay cell viability, assay kinetics of cell cycle phases, annexin V assay/ propidium iodide, test of mitochondrial membrane potential, test comet and gene expression through real time PCR. Both antiproliferative and cytotoxic activity revealed that K562 cells cultured with ruthenium (III) compound showed meaningful decrease in proliferation. Ruthenium(III) compound induced the IC50 value was of 18.28 μM set againts the cell cycle profiles cells not treated. Flow cytometric analysis indicated a sub-G1 arresting effect of ruthenium compound on K562 cells. Through the cell viability assay through MTT reduction technique, it was found that the complex of ruthenium (II) phosphine coordinated and nitrile presented cytotoxic activity when facing the tumor strain S180 with IC50 17.02±8.21μM and IC50 de 53.73 ± 5.71 μM for lymphocyte. When analyzing the cell cycle of tumor cells S180 treated with complex of ruthenium (II) caused increase in cells in G0/G1 and in S phase decreased. We observed an increase G2 / M. In the analysis of apoptosis assays, the results pointed that the complex ruthenium (II) induced cell death via apoptosis in tumor strain S180 as proved the increase in annexin cells V positive, depolarization of the mitochondrial membrane potential, activation of caspase 3 (Casp3) and 8 (Casp8) and increased expression levels of caspase-3 (Casp3) (mRNA), Bax (mRNA) and Tp53. The results lead to the conclusion that both complexes of ruthenium (II) and (III) induce cytotoxic activity against cell models tested, and that this activity correlates with alterations in cell cycle phases and induction of cell death via apoptosis. |
publishDate |
2014 |
dc.date.issued.fl_str_mv |
2014-03-19 |
dc.date.accessioned.fl_str_mv |
2015-10-27T14:38:45Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
PEREIRA, F. C. Mecanismo de morte celular induzida por complexos de rutênio II e III em diferentes linhagens tumorais. 2014. 162 f. Tese (Doutorado em Biologia) - Universidade Federal de Goiás, Goiânia, 2014. |
dc.identifier.uri.fl_str_mv |
http://repositorio.bc.ufg.br/tede/handle/tede/4787 |
dc.identifier.dark.fl_str_mv |
ark:/38995/0013000006d5g |
identifier_str_mv |
PEREIRA, F. C. Mecanismo de morte celular induzida por complexos de rutênio II e III em diferentes linhagens tumorais. 2014. 162 f. Tese (Doutorado em Biologia) - Universidade Federal de Goiás, Goiânia, 2014. ark:/38995/0013000006d5g |
url |
http://repositorio.bc.ufg.br/tede/handle/tede/4787 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.program.fl_str_mv |
6883982777473437920 |
dc.relation.confidence.fl_str_mv |
600 600 600 600 600 |
dc.relation.department.fl_str_mv |
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