RESUMO
It has been reported that the antitumor drug doxorubicin (Dox) exerts its toxic effects via GATA-4 depletion and that over-expression of GATA-4 reverses Dox-induced toxicity and apoptosis; however, the precise mechanisms remain unclear. In this study, we observed, for the first time, that EGF protects cells against Dox-mediated growth arrest, G2/M-phase arrest, and apoptosis. Additionally, EGF expression was down-regulated in Dox-treated cells and up-regulated in GATA-4 over-expressing cells. Utilizing real-time PCR and western blotting analysis, we found that the expression of the cell cycle-associated protein cyclin D1 was inhibited in GATA-4-silenced cells and Dox-treated cells and was enhanced in GATA-4 over-expressing cells and EGF-treated cells. Furthermore, EGF treatment reversed the inhibited expression of cyclin D1 that was mediated by GATA-4 RNAi or Dox. Our results indicate that EGF, as a downstream target of Dox, may be involved in Dox-induced toxicity as well as in the protective role of GATA-4 against toxicity induced by Dox via regulating cyclin D1 expression, which elucidates a new molecular mechanism of Dox toxicity with important clinical implications.
Assuntos
Antibióticos Antineoplásicos/farmacologia , Ciclina D1/metabolismo , Doxorrubicina/farmacologia , Fator de Crescimento Epidérmico/farmacologia , Fator de Transcrição GATA4/metabolismo , Animais , Apoptose , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Fator de Transcrição GATA4/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Células HEK293 , Humanos , CamundongosRESUMO
The regulation of cardiac differentiation is critical for maintaining normal cardiac development and function. The precise mechanisms whereby cardiac differentiation is regulated remain uncertain. Here, we have identified a GATA-4 target, EGF, which is essential for cardiogenesis and regulates cardiac differentiation in a dose- and time-dependent manner. Moreover, EGF demonstrates functional interaction with GATA-4 in inducing the cardiac differentiation of P19CL6 cells in a time- and dose-dependent manner. Biochemically, GATA-4 forms a complex with STAT3 to bind to the EGF promoter in response to EGF stimulation and cooperatively activate the EGF promoter. Functionally, the cooperation during EGF activation results in the subsequent activation of cyclin D1 expression, which partly accounts for the lack of additional induction of cardiac differentiation by the GATA-4/STAT3 complex. Thus, we propose a model in which the regulatory cascade of cardiac differentiation involves GATA-4, EGF, and cyclin D1.
Assuntos
Diferenciação Celular/fisiologia , Fator de Crescimento Epidérmico/metabolismo , Fator de Transcrição GATA4/metabolismo , Coração/embriologia , Modelos Biológicos , Miocárdio/citologia , Transdução de Sinais/fisiologia , Animais , Western Blotting , Linhagem Celular Tumoral , Imunoprecipitação da Cromatina , Técnicas Histológicas , Imunoprecipitação , Camundongos , Reação em Cadeia da Polimerase em Tempo Real , Fatores de TempoRESUMO
The iron-pillared bentonite (Fe-Ben) was prepared by ion-exchange using the natural bentonite (GZ-Ben) from Gaozhou, China, at room temperature without calcination. Both Fe-Ben and GZ-Ben were characterized by X-ray diffraction, N(2) adsorption and Fourier transform infrared spectroscopy. The results show that the d(001) value and surface area of the bentonite material increased after iron pillaring. Fe-Ben adsorbed much more Rhodamine B (RhB) than GZ-Ben, which can be ascribed to the special surface properties and large surface area of Fe-Ben. The optimum pH value for the adsorption of RhB on Fe-Ben is 5.0. The adsorption of RhB onto Fe-Ben can be well described by the pseudo-second-order kinetic model and the intraparticle diffusion kinetic model. The adsorption isotherm of RhB onto Fe-Ben matches well with the Langmuir model.