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Previous issue date: 2017-03-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The redox-active chemical properties of trace element copper make it an essential cofactor for several cellular mechanisms, as for ROS production. Polyphenols have been used in the pro-oxidant mechanism of action of interaction with endogenous Cu (II) ions for the production of reactive oxygen species in excess as a treatment for malignancies, leading to apoptosis. This work studied Cu (II) ions in the complex formation mechanism with Resveratrol in MCF-7 cells model. We analyzed the selectivity of Resveratrol in relation to the cupric metal ion using HPLC (High-Performance Liquid Chromatography) and the formation of the compound Resveratrol-Copper through UV-VIS (Ultra-Violet Visible Spectrophotometry). We analyzed the cellular morphology and location of the metal ion by MET-EDS (Electronic Transmission microscopy with Dispersive X-ray Spectroscopy). Cell death by apoptosis and quantification of ROS in said cell line with copper enrichment and treated with Resveratrol was made by flow cytometry. The results show the selectivity of the polyphenol compound by copper ion as well as the formation of the complex Resveratrol-Copper in extracellular conditions, however even with verification of endogenous copper accumulation in physiological conditions in vitro, the formation of that complex did not occur because there was no production of ROS and therefore, no cell death. In short, our research reveals that for in vitro conditions for the MCF-7 line, there's no Resveratrol-Copper complex formation as observed in sub-lethal quantities of chemically enriched cells with CuSO4 and treated with Resveratrol. / As propriedades qu?micas redox-ativas do oligoelemento cobre o tornam cofator essencial para diversos mecanismos celulares como o de produ??o de ROS. Polifen?is v?m sendo utilizados no mecanismo de a??o pr?-oxidante de intera??o com ?ons Cu (II) end?geno para a produ??o dessas esp?cies oxig?nio reativas em excesso como tratamento de malignidades, levando a apoptose. O presente trabalho estudou os ?ons Cu (II) no mecanismo de forma??o do complexo com Resveratrol em modelo de c?lulas MCF-7. Analisamos a seletividade do Resveratrol frente ao ?on met?lico c?prico utilizando CLAE (Cromatografia L?quida de Alta Efici?ncia) e verificou-se a forma??o do complexo Resveratrol-Cobre por meio de UV-VIS (Espectrofotometria de Ultra-Violeta Vis?vel). Analisou-se a morfologia celular e localiza??o do ?on met?lico por MET-EDS (Microscopia de Transmiss?o Eletr?nica com Espectroscopia Dispersiva de Raios-X). Morte celular por apoptose e quantifica??o de ROS na linhagem enriquecida com cobre e tratadas com Resveratrol foi feita por Citometria de Fluxo. Os resultados mostram a seletividade do composto polifen?lico pelo ?on cobre bem como a forma??o do complexo Resveratrol-Cobre em condi??es extracelulares, por?m mesmo com a verifica??o de ac?mulo de cobre end?geno, em condi??es fisiol?gicas in vitro n?o foi constatada a forma??o do referido, pois n?o houve forma??o de ROS e morte celular conseguinte. Em suma, Nossa pesquisa revela que em condi??es in vitro para a linhagem MCF-7, n?o h? forma??o de complexo Resveratrol-Cobre como observado quimicamente em quantidades sub-letais de c?lulas enriquecidas com CuSO4 e tratadas com Resveratrol.
| Identifer | oai:union.ndltd.org:IBICT/oai:tede2.pucrs.br:tede/7590 |
| Date | 27 March 2017 |
| Creators | Volkart, Priscylla Andrade |
| Contributors | Souto, Andr? Arigony, Vargas, Jos? Eduardo |
| Publisher | Pontif?cia Universidade Cat?lica do Rio Grande do Sul, Programa de P?s-Gradua??o em Biologia Celular e Molecular, PUCRS, Brasil, Faculdade de Bioci?ncias |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da PUC_RS, instname:Pontifícia Universidade Católica do Rio Grande do Sul, instacron:PUC_RS |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | 8198246930096637360, 500, 500, 500, 600, 36528317262667714, -1634559385931244697, 2075167498588264571 |
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