RESUMEN

When F9 teratocarcinoma cells are treated with retinoic acid plus cyclic AMP (RACF9) they differentiate into parietal endoderm. Differentiation is accompanied by the acquisition of substrate adhesion sites and a change in the pattern of gene expression, including the synthesis of tissue-type plasminogen activator (tPA). We demonstrate here that dihydrocytochalasin B (DHCB) treatment of differentiating F9 cells prevents the assembly of a structured actin cytoskeleton and generates a more rounded and stellate cell morphology. This morphological change is accompanied by the accumulation of the usually visceral endoderm-specific marker urokinase-type plasminogen activator (uPA) and an increase in tPA levels in comparison to untreated RACF9 controls. The increase in tPA accumulation is preceded by an increase in tPA mRNA levels. These effects are reversible, with a lag, when DHCB is removed, and PA accumulation can be stimulated within 24 h when differentiated cells are exposed to DHCB. Exposure to the microtubule disrupting agent colchicine has no effect on uPA or tPA accumulation. In addition, antibody directed against the beta 1 integrin subunit can also specifically elicit increased PA production. Thus disturbing the cytoskeleton and cytoskeleton associated substrate adhesions promotes PA accumulation.

Asunto(s)

Citoesqueleto/fisiología , Endodermo/patología , Matriz Extracelular/fisiología , Activadores Plasminogénicos/metabolismo , Actinas/química , Actinas/metabolismo , Animales , Colchicina/farmacología , AMP Cíclico/farmacología , Citocalasina B/análogos & derivados , Citocalasina B/farmacología , Citoesqueleto/química , Citoesqueleto/metabolismo , Electroforesis en Gel de Poliacrilamida , Endodermo/química , Endodermo/metabolismo , Matriz Extracelular/química , Matriz Extracelular/metabolismo , Técnica del Anticuerpo Fluorescente , Expresión Génica , Sueros Inmunes/inmunología , Integrinas/inmunología , Activadores Plasminogénicos/análisis , Activadores Plasminogénicos/genética , ARN Mensajero/análisis , ARN Mensajero/genética , Teratoma , Factores de Tiempo , Activador de Tejido Plasminógeno/análisis , Activador de Tejido Plasminógeno/genética , Activador de Tejido Plasminógeno/metabolismo , Tretinoina/farmacología , Células Tumorales Cultivadas , Activador de Plasminógeno de Tipo Uroquinasa/análisis , Activador de Plasminógeno de Tipo Uroquinasa/genética , Activador de Plasminógeno de Tipo Uroquinasa/metabolismo