The effect of 12-O-tetradecanoylphorbol-13-acetate and other tumor promoters on the colony formation of rat tracheal epithelial cells in culture.

Abstract

Normal primary rat tracheal epithelial (RTE) cells were isolated and exposed in culture to tumor promoters and irritants of diverse chemical classes. The phorbol derivative class of tumor promoters greatly stimulated colony forming efficiency (CFE) in culture. The efficacies of the agents tested were ranked in the order: mezerein greater than 12-O-tetradecanoyl-phorbol-13-acetate (TPA) greater than phorbol didecanoate greater than phorbol dibutyrate greater than phorbol dibenzoate greater than 4-O-methyl TPA greater than phorbol diacetate. The parent alcohol phorbol did not stimulate CFE under the conditions tested. The indole alkaloid tumor promoter teleocidin stimulated CFE at concentrations at least 10-fold lower than those required for TPA. Other irritants and non-phorbol ester tumor promoters such as anthralin, benzoyl peroxide, calcium ionophore A23187, and ethylphenyl propiolate were either inactive or reduced CFE. Phenobarbital marginally stimulated CFE at one concentration but reduced CFE at higher concentrations. Increases in CFE elicited by TPA and analogs were dependent upon the time of addition of TPA to the cultures. Maximum increases in CFE were observed when the cells were plated into medium containing TPA. If TPA was added 40 h after plating, stimulation of CFE did not occur. This 40 h time interval may represent a crucial period for the commitment of RTE stem cells to proliferation or differentiation. Whether the stimulation of colony formation seen in normal RTE cells exposed to phorbol derivatives also occurs in carcinogen-altered cells, thereby causing their proliferative expansion, remains to be determined.