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Master's Dissertation
Full name
Elaine Silveira
Knowledge Area
Date of Defense
São Paulo, 2014
Lotfi, Claudimara Ferini Pacicco (President)
Bezerra Neto, João Evangelista
Freitas, Vanessa Morais
Title in Portuguese
Análise da viabilidade das células H295R, linhagem de carcinoma adrenocortical tumoral humano, tratadas com diferentes drogas antitumorais.
Keywords in Portuguese
Carcinoma adrenocortical
Abstract in Portuguese
O carcinoma adrenocortical (ACC) é um tumor maligno raro. O objetivo foi testar a ação de drogas antitumorais na linhagem H295R, em cultura 2D e 3D, analisadas por MTS, ensaios multiparamétricos, e microscopia confocal. Em monocamada (2D), as drogas tiveram maior efeito quando utilizadas com o mitotano 10 mM, sendo: everolimus 10 mM (50±0,02% p ≤ 0.001), imatinib (10 mM 52±0.01% p ≤ 0.001), sunitinib 5 mM (47±0.02% p ≤ 0.001), e nilotinib 5 mM (59±0,03% p ≤ 0.001), com evidência de apoptose. Nos esferoides (3D) foi necessário mitotano 30 M com everolimus 10 mM para se obter diminuição de 32±0.02% p ≤ 0.001 na viabilidade das células, e com nilotinib 10 mM para redução de 57±0.03% p ≤ 0.001, com evidência de necrose e apoptose. Em resumo, os dados sugerem que os esferoides são mais resistentes aos tratamentos, como ocorre com tumores in vivo, e podem representar uma importante abordagem para estudos de ACC. O nilotinib foi o que induziu as melhores respostas, tanto no modelo em 2D quanto 3D, resultados que se apresentam promissores para o tratamento dos carcinomas adrenocorticais.
Title in English
Antitumor effects of different cytotoxic drugs on the human adrenocortical tumor cells H295R.
Keywords in English
Adrenocortical carcinoma
Abstract in English
Adrenocortical carcinoma (ACC) is a rare malignant tumor. The objective was to test the antitumor drugs action in the H295R cell line, in 2D and 3D culture system, by using MTS, multiparameter assay (High Content Screening) and confocal microscopy. In monolayer, all drugs were more effective in combination with mitotane 10 mM: everolimus 10 mM (50±0.02%), 10 mM imatinib (52±0.01%), 5 mM sunitinib (47±0.02%), and 5 mM nilotinib (59±0.03%). The spheroids required mitotano 30 mM with everolimus 10 mM to decrease cell viability (32±0.02% p ≤ 0.001); and with 10 mM nilotinib to inhibit cell viability in 57±0.03% (p ≤ 0.001), with induction of apoptosis and necrosis. In summary, the spheroids were more resistant to treatment and may represent an important approach for studies of ACC. Nilotinib was the tyrosine kinase inhibitor that, either alone or in combination with mitotane, induced higher cytotoxicity, in both 2D and 3D cell cultures, showing promising results for the treatment of adrenocortical carcinoma, as well as for the studies of its mechanism of action.
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