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1.
Exp Dermatol ; 28(9): 1079-1082, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31338879

RESUMO

Since Drosophila melanogaster has proven to be a useful model system to study phenotypes of oncogenic mutations and to identify new anti-cancer drugs, we generated human BRAFV600E homologous dRaf mutant (dRafA572E ) Drosophila melanogaster strains to use these for characterisation of mutant phenotypes and exploit these phenotypes for drug testing. For mutant gene expression, the GAL4/UAS expression system was used. dRafA572E was expressed tissue-specific in the eye, epidermis, heart, wings, secretory glands and in the whole animal. Expression of dRaf A572E under the control of an eye-specific driver led to semi-lethality and a rough eye phenotype. The vast majority of other tissue-specific and ubiquitous drivers led to a lethal phenotype only. The rough eye phenotype was used to test BRAF inhibitor vemurafenib and MEK1/2 inhibitor cobimetinib. There was no phenotype rescue by this treatment. However, a significant rescue of the lethal phenotype was observed under a gut-specific driver. Here, MEK1/2 inhibitor cobimetinib rescued Drosophila larvae to reach pupal stage in 37% of cases as compared to 1% in control experiments. Taken together, the BRAFV600E homolog dRaf A572E exerts mostly lethal effects in Drosophila. Gut-specific dRaf A572E expression might in future be developed further for drug testing.


Assuntos
Azetidinas/farmacologia , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , MAP Quinase Quinase Quinases/antagonistas & inibidores , Piperidinas/farmacologia , Proteínas Proto-Oncogênicas c-raf/genética , Animais , Proteínas de Drosophila/biossíntese , Proteínas de Drosophila/deficiência , Proteínas de Drosophila/fisiologia , Avaliação Pré-Clínica de Medicamentos , Regulação da Expressão Gênica no Desenvolvimento , Genes Letais , Intestinos/enzimologia , Larva , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Especificidade de Órgãos , Fenótipo , Inibidores de Proteínas Quinases/farmacologia , Proteínas Proto-Oncogênicas B-raf/fisiologia , Proteínas Proto-Oncogênicas c-raf/biossíntese , Proteínas Proto-Oncogênicas c-raf/deficiência , Proteínas Proto-Oncogênicas c-raf/fisiologia , Vemurafenib/farmacologia
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