Efeito do meio condicionado em c??lulas pr??-tratadas com extrato hidroalco??lico de guaran?? em c??lulas tumorais

Autor: Souza, Patr??cia Rayn?? Simas de
Přispěvatelé: Faria, Jerusa Ara??jo Quint??o Arantes, Gomes, Dawidson Assis, Sadahiro, Aya
Jazyk: portugalština
Rok vydání: 2021
Předmět:
Zdroj: Biblioteca Digital de Teses e Dissertações da UFAM
Universidade Federal do Amazonas (UFAM)
instacron:UFAM
Popis: Submitted by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2021-08-26T19:02:05Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) ITEM INDISPON??VEL - Solicite c??pia.pdf: 84143 bytes, checksum: bec574cfc2dc76cf8ba0e289d73276b9 (MD5) Approved for entry into archive by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2021-08-27T00:44:28Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) ITEM INDISPON??VEL - Solicite c??pia.pdf: 84143 bytes, checksum: bec574cfc2dc76cf8ba0e289d73276b9 (MD5) Made available in DSpace on 2021-08-27T00:44:28Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) ITEM INDISPON??VEL - Solicite c??pia.pdf: 84143 bytes, checksum: bec574cfc2dc76cf8ba0e289d73276b9 (MD5) Previous issue date: 2021-08-20 CAPES - Coordena????o de Aperfei??oamento de Pessoal de N??vel Superior The tumor microenvironment is a determinant in the programming, regression or progression of the tumor, as it has the ability to modulate tumor cells through the release of soluble factors in the environment. The study of the antineoplastic effect of drugs, as well as that of herbal medicines, encompasses the action of different cell types and how the action of these compounds modulate these cells present in the tumor microenvironment. In addition, in the scenario of herbal medicines with antineoplastic action, guarana (Paullinia cupana) has emerged as a candidate. Objective: The objective of this work was to evaluate the direct effect of the hydroalcoholic extract of guarana (EHG) on different cell lines and the effect of conditioned media (MCs) on macrophages and normal epithelial cells pretreated with EHG on the proliferative and migratory profile of breast and lung tumor cells. Materials and Methods: The values of the minimum inhibitory concentration of EHG, as well as the antioxidant activity and evaluation of the death profile were conducted in cell lines. Conditioned media were generated in cells pretreated with the highest non-cytotoxic concentration of EHG. The effects of secretoma were evaluated in tumor cells by means of the clonogenicity test, the assessment of viability, cell migration and analysis of the cell cycle profile. Results: The results showed that the values obtained from direct treatment with EHG were heterogeneous among the different cell lines evaluated: MDA-MB231, MCF7, MCF10A, A549, BEAS-2B, NIH- 3T3, MRC5, NCTC clone 929, differentiated THP-1 in macrophage. The most resistant strain was lung adenocarcinoma (IC50=17.652??10.22 ??g/ml), while the most sensitive was the murine fibroblast (IC50=1.504??34.78 ??g/ml). EHG demonstrated antioxidant activity by reducing the levels of reactive oxygen species (ROS) in the MDA-MB 231 strain, both at baseline and under oxidative stress conditions. Direct treatment of EHG for 24h with the IC50 value (12,022??10.73 ??g/ml) directs BEAS-2B cells to an apoptotic death profile. The results showed that the MCs in MCF10A were able to reduce the viability and clonogenic capacity of A549, while those in MCF7 only reduced the clonogenic capacity. The MCF10A MC CTL increased the amount of A549 cell cycle G1 phase cells. MC from EHG-pretreated lung epithelial cells increases the wound closure capacity of A549 cells while reducing clonogenic capacity. The MCF7 lineage when treated with secretoma of differentiated cells into macrophages reduces viability within 72 hours of treatment, but the group that was stimulated with LPS restores cell viability, these data are not visualized in the clonogenic assay where all groups reduce clonogenic capacity. A549 is responsive only to groups that are stimulated with LPS in the clonogenic, increasing the percentage of migration of cells that were treated with secretoma of differentiated cells into macrophages pretreated with EHG and stimulated with LPS, while stimulating the formation of vacuoles in the tumor cells and changes morphology to fusiform, losing cell-cell adhesion. Conclusion: MCs act in different ways in the proliferative capacities of the studied tumor lines, however it is observed that the secretoma of cells pretreated with EHG increased the migratory capacity of A549. Future experiments are needed to elucidate the mechanism of action of the EHG in the modulation of macrophage secretama, as well as to evaluate the induction of the epithelial-mesenchymal transition process in tumor cells treated with the conditioned medium. O microambiente tumoral ?? um determinante na regress??o ou progress??o do tumor apresentando a capacidade de modular as c??lulas tumorais. O estudo do efeito antineopl??sico de quimioter??picos, abrange a modula????o das c??lulas presentes no microambiente do tumor. Em adi????o, no cen??rio de fitoter??picos com a????o antineopl??sica, o guaran?? (Paullinia cupana) tem emergido como um candidato promissor. Objetivo: O objetivo deste trabalho consistiu em avaliar o efeito direito do extrato hidroalco??lico de guaran?? (EHG) em diferentes linhagens celulares e o efeito dos meios condicionados (MCs) em macr??fagos e em c??lulas epiteliais normais pr??-tratadas com EHG no perfil proliferativo e migrat??rio de c??lulas tumorais de mama e de pulm??o. Materiais e M??todos: Os valores da concentra????o inibit??ria m??nima do EHG, bem como a atividade antioxidante e avalia????o do perfil de morte foram conduzidos em linhagens celulares. Os meios condicionados foram gerados nas c??lulas pr??-tratadas com a maior concentra????o n??o citot??xica de EHG. Os efeitos do secretoma foram avaliados nas c??lulas tumorais pelos ensaios de viabilidade, clonogenicidade, migra????o celular e an??lise do perfil do ciclo celular. Resultados: A caracteriza????o por RP-HLPC identificou a presen??a das metilxantinas cafe??na, teobromina e teofilina, bem como catequina no EHG. O perfil de viabilidade e os valores de CI50 foram heterog??neos entre as distintas linhagens celulares: MDA-MB231, MCF7, MCF10A, A549, BEAS-2B, NIH- 3T3, MRC5, NCTC clone 929, THP-1 diferenciada em macr??fago. A linhagem mais resistente A549 adenocarcinoma de pulm??o (CI50=17.652??10.22 ??g/ml), enquanto a mais sens??vel foi o fibroblasto murino (CI50=1.504??34.78 ??g/ml). Tratamento com EHG reduziu os n??veis de esp??cies reativas de oxig??nio (EROs) na linhagem MDA- MB 231 e em THP-1 diferenciada em macr??fagos em condi????o de estresse oxidativo. O tratamento direto de EHG direciona c??lulas BEAS-2B para um perfil de morte por apoptose. Os meios condicionados (MCs) gerados em MCF10A tratada ou n??o com EHG reduziram a viabilidade e a capacidade clonog??nica, elevando o n??mero de c??lulas tumorais A549 na fase G1 do ciclo celular. Ademais, estes secretomas promoveram a redu????o da capacidade clonog??nica das c??lulas tumorais de mama MCF-7. O MC gerado em BEAS-2B previamente tratadas com EHG aumentou a capacidade de fechamento de ferida de A549 com redu????o da sua capacidade clonog??nica. O secretoma de c??lulas THP-1 diferenciadas em macr??fagos reduz a viabilidade de MCF-7, ao passo que o MC gerado em macr??fagos desafiados com LPS ?? capaz de restaurar a viabilidade. C??lulas A549 aumentam o percentual de migra????o quando incubadas com o secretoma de c??lulas diferenciadas em macr??fagos pr??-tratadas com EHG e estimulado com LPS. Adicionalmente, observa-se altera????o morfol??gica de tais c??lulas tumorais com a forma????o de vac??olos, morfologia fusiforme e perda da ades??o c??lula-c??lula. Conclus??o: Os MCs gerado em BEAS-2B, MCF10A e THp-1 diferenciadas em macr??fagos agem de modo distinto na capacidade proliferativa das linhagens tumorais estudadas. No entanto, observa-se que o secretoma de c??lulas pr??- tratadas com EHG aumentaram a capacidade migrat??ria de A549. Experimentos futuros s??o necess??rios para elucidar o mecanismo de EHG na modula????o do secretoma de macr??fagos, bem como avaliar a indu????o do processo de transi????o epit??lio-mesenquimal nas c??lulas tumorais tratadas com o meio condicionado.
Databáze: OpenAIRE
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