Transient expression of the transformed phenotype stimulated by 12-O-tetradecanoyl phorbol-13-acetate

Autor: H.K. Plummer, Carol A. Heckman
Rok vydání: 1990
Předmět:
Zdroj: Experimental Cell Research. 188:66-74
ISSN: 0014-4827
DOI: 10.1016/0014-4827(90)90279-j
Popis: The objective of the present work was to determine whether a short-term perturbation of cells by 12-O-tetradecanoyl phorbol-13-acetate (TPA) treatment caused shape changes identical to those found in oncogenic transformation. A cell line derived from the rat respiratory tract epithelium, 1000 W, was used, in which shape changes had been identified previously as the cells underwent transformation during long-term growth in vitro. These changes corresponded to a steeper rise of the cell from the substrate and a smoothing of the surface contours throughout the periphery of the cell. The phenotype was measured by maximum likelihood estimation, based on the values of several geometrical shape descriptors. With continuous TPA treatment, the cells adopted a transformed phenotype by 2 h. The effect was maximal by 5 h but began to decline by 10 h. Shape change in the opposite direction was stimulated by treatment with the protein kinase C inhibitor staurosporine, and its effects were counteracted if the cells were simultaneously exposed to TPA. No appreciable metabolism of [20-3H]TPA occurred until 24 h after treatment. Enumerating the shape descriptors whose values composed the transformed phenotype indicated that the TPA-stimulated changes were qualitatively similar to those accompanying oncogenic transformation. The subsequent alterations, however, involved few of the variables that composed the transformed phenotype and therefore did not represent a true reversal of the change. Changes observed up to 5 h were not dependent on new RNA synthesis but required continued protein synthesis.
Databáze: OpenAIRE