RESUMO
The in vitro phosphorylated and non-phosphorylated Hsp27 forms concentrations and Bcl-2 proteins affected by Hsp27 inhibition were studied in Jurkat-line tumor cells and healthy donor mononuclear lymphocytes by Western blotting technique. The Hsp27 inhibition causes the increase of intracellular Bax protein concentration and the decrease of Bcl-2 level leading to an increase of apoptotic changes in Jurkat line cells.
Assuntos
Proteínas Reguladoras de Apoptose/metabolismo , Apoptose , Proteínas de Choque Térmico HSP27/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteína X Associada a bcl-2/metabolismo , Proteína de Morte Celular Associada a bcl/metabolismo , Apoptose/efeitos dos fármacos , Western Blotting , Humanos , Células Jurkat/efeitos dos fármacos , Células Jurkat/metabolismo , Chaperonas Moleculares/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/efeitos dos fármacos , Proteína X Associada a bcl-2/efeitos dos fármacos , Proteína de Morte Celular Associada a bcl/efeitos dos fármacosRESUMO
The role of Hsp27 (heat shock protein 27) chaperone in regulation of THP-1 tumor cell apoptosis was studied. Realization of tumor cell apoptosis under conditions of in vitro culturing with Hsp27 specific inhibitor (KRIBB3) was evaluated by fluorescent microscopy with FITC-labeled annexin V and propidium iodide. Measurements of Bcl-2 family proteins (Bcl-2, Bax, Bad) in tumor cells incubated with Hsp27 inhibitor were carried out by Western blotting. Chaperone Hsp27 acted as apoptosis inhibitor in THP-1 tumor cells modulating the proportion of antiapoptotic (Bcl-2) and proapoptotic (Bax and Bad) proteins.
Assuntos
Anisóis/farmacologia , Apoptose/fisiologia , Proteínas de Choque Térmico HSP27/metabolismo , Isoxazóis/farmacologia , Leucemia/patologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Adolescente , Adulto , Anexina A5/metabolismo , Proteínas Reguladoras de Apoptose , Linhagem Celular Tumoral , Feminino , Proteínas de Choque Térmico HSP27/antagonistas & inibidores , Humanos , Leucócitos Mononucleares , Masculino , Pessoa de Meia-Idade , Chaperonas Moleculares/metabolismo , Células Precursoras de Monócitos e Macrófagos/patologia , Adulto JovemRESUMO
Programmed death of Jurkat tumor cells was studied under conditions of culturing with 17-AAG selective inhibitor of heat shock protein with a molecular weight of 90 kDa and etoposide. Apoptosis realization was evaluated by fluorescent microscopy with FITC-labeled annexin V and propidium iodide. Activity of caspase-3 was evaluated spectrophotometrically. Inhibition of heat shock protein with a molecular weight of 90 kDa activated the apoptotic program in Jurkat tumor cells and etoposide-induced apoptosis. The heat shock protein with a molecular weight of 90 kDa acted as apoptosis inhibitor in tumor cells.
