Cellular interactions of phorbol myristate acetate in tumor promotion.

Abstract The radioactivity from tritium-labeled phorbol myristate acetate is bound to mouse skin with a half-life of approx. 24 h. Autoradiographs revealed that most of the label was associated with the kerato-sebaceous layer over the squamous epithelial layer. The major portion of the radioactivity was recovered in the lipid-containing fractions of a Schmidt-Thannhauser extraction. In cell cultures of 3T3 mouse embryo fibroblasts, maximal binding of [3H]PMA was achieved in 18 h and was stable for up to 4 days in the presence of the labeled tumor-promoting agent. Autoradiographs of formalin-fixed cells showed extensive cytoplasmic labeling with no substantial nuclear activity. Methanol fixation resulted in complete removal of the tritium. Sedimentation analysis in Dextran gradients of subcellular fractions of 3T3 cells exposed to [3H]PMA demonstrated that the bulk of bound label was associated with a Na+-K+ ATPase-rich protein peak which has the properties of cell membranes.

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