Metabolism and macromolecular covalent binding of benzo[a]pyrene in cultured Fischer-344 rat lung type II epithelial cells

Pulmonary biotransformation of many xenobiotics may be important for the mutagenic, carcinogenic and/or toxic response of lung tissue to these compounds. Recently, a lung epithelial cell line (designated LEC), with morphological characteristics suggestive of type II cell origin, was developed in our laboratory. When LEC cells were co-cultivated with Chinese hamster ovary (CHO) cells in a cell-mediated mutagenesis assay, LEC metabolized promutagens to metabolites mutagenic to the CHO cells [A. P. Li, A. L. Brooks, J. M. Benson and F. F. Hahn, Environ. Mutagen. 4, 407 (1982)]. In the present investigation, rates of benzo[a]pyrene (BaP) metabolism in type II lung cells were determined, and the effects of various pollutants on rates of BaP metabolism and covalent binding of BaP to LEC macromolecules were measured. Cultures of LEC cells were incubated for 24 hr with 5 microM [14C]BaP, and the culture medium was analyzed for organic- and water-soluble metabolites. LEC cells metabolized BaP to BaP-7,8-diol and BaP-9,10-diol with total rates of formation of these metabolites measured at 500-600 pmoles per 10(6) cells per 24 hr. BaP-9,10-diol was the major metabolite accounting for about 80% of the total BaP metabolized. Enzyme hydrolysis studies revealed the presence of small quantities (less than 20% of BaP metabolized) of the glucuronide conjugates of BaP-7, 8-diol and 9-hydroxy-BaP. Pretreatment of LEC cells with benz[a]anthracene, coal gas condensate, or diesel exhaust particle extract (DEP) prior to incubation with BaP resulted in a 2- to 5-fold increase in overall rates of BaP metabolism. The largest increase in covalent binding of [14C]BaP equivalents to LEC macromolecules was seen after LEC cells were pretreated with DEP (3-fold). The data suggest that lung epithelial cells may play an important role in the biological fate of inhaled xenobiotics.