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Master's Dissertation
DOI
10.11606/D.5.2016.tde-11042016-113530
Document
Author
Full name
Mariana Daou Verenhitach
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2015
Supervisor
Committee
Maria, Durvanei Augusto (President)
Matera, Julia Maria
Gomes, Ligia Ferreira
Title in Portuguese
Avaliação dos efeitos antitumorais e antiproliferativos do peptídeo INKKI em células de adenocarcinoma de mama MCF-7
Keywords in Portuguese
Apoptose
Caseínas
Caspases
Hidrólise
INKKI
Neoplasias da Mama
Peptídeos
Abstract in Portuguese
O INKKI é um peptídeo catiônico anfipático derivado da ?-caseina bovina, análogo ao Mastoparan, isolado do veneno da vespa Vespula lewisi. Diferentes estudos mostraram sua atividade antiproliferativa e antitumoral in vitro com citotoxidade seletiva em linhagens de células de melanoma B16F10 em relação às células normais. Em estudos In vivo, INKKI demonstrou potente ação estimuladora da atividade fagocitária de macrófagos peritoneais. Quando aplicado diretamente em tumores, reduziu a massa tumoral, diminuindo o tempo de progressão do tumor e a formação de metástases, aumentando a probabilidade de sobrevida. Neste trabalho foram investigados os efeitos in vitro do peptídeo INKKI em linhagens de células de adenocarcinoma de mama e em células normais. As linhagens de células tumorais de mama utilizadas foram MDA-231, MCF-7 e T47D humanas, mama murina de Ehrlich e, como células normais, os fibroblastos e as células endoteliais. Foram avaliadas a atividade citotóxica, a viabilidade celular, a alteração do potencial elétrico mitocondrial, os efeitos na progressão e parada do ciclo celular e a investigação da morte por apoptose. Os resultados revelaram que o peptídeo INKKI apresenta citotoxidade seletiva dose-dependente nas células tumorais, modulando negativamente o potencial elétrico mitocondrial. O índice de proliferação celular das células tratadas diminuiu, com a parada do ciclo celular na fase G0/G1. O peptídeo induziu a morte celular por apoptose, a qual demonstrou ocorrer dependente da via mitocondrial e de forma caspaseindependente. O peptídeo INKKI é um potente modulador das atividades antiproliferativa e antitumoral da linhagem de células de adenocarcinoma de mama humana MCF-7
Title in English
Evaluation of antitumor and antiproliferative effects of the INKKI peptide in breast adenocarcinoma MCF-7 cell line
Keywords in English
Apoptosis
Breast neoplasms
Caseins
Caspases
Hidrolysis
INKKI
Peptide
Abstract in English
The INKKI is derived from the bovine hidrolisis of ?-casein, a cationic and amphipathic peptide, analogous to mastoparan peptide isolated from the venom of the wasp Vespula lewisi. In in vitro studies, INKKI showed selective cytotoxicity in melanoma tumor cell B16F10 in relation to normal cells. In the in vivo tests, INKKI peptide has been shown to be a potent stimulator of the phagocytic activity of treated macrophages and, when applied directly into tumors, reduced tumor mass, decreasing the time progression and metastasis formation, increasing the probability of survival. This work investigated the in vitro effects of tumor peptide INKKI in cell line of human breast adenocarcinoma MCF-7. The tumor cells used were lineages of human breast tumor cells MCF7, MDA-231 and T47D and murine tumor cells (Ehrlich). The normal cells used were fibroblasts and endothelial cells. The study evaluated the cytotoxic activity, cell viability, change of mitochondrial electrical potential, effects on the progression and arrest of the cell cycle and investigation of induced death by apoptosis. The results revealed that peptide presents selective dose-dependent cytotoxicity in tumor cells by modulating negatively the mitochondrial electric potential. The index of cell proliferation of cells treated declined, with the induction of cell cycle arrest at phase G0/G1. The peptide-induced cell death by apoptosis was demonstrated to occur via mitochondrial pathway and in a caspase-independent manner. The INKKI peptide is a potent modulator of antiproliferative and antitumor activities of the breast adenocarcinoma cell line
 
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Publishing Date
2016-04-11
 
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