Syrian hamster embryo cells were treated with galactose oxidase, followed by reduction with tritiated sodium borohydride at pH 7.4. The labeling patterns of galactosyl and N-acetyl galactosaminyl residues on the cell surface were altered in comparing scraped vs. unscraped and buffer vs. media-soaked cells treated with galactose oxidase. From these preliminary studies, the procedure to be used in most of the asbestos treatment studies was to treat cells in situ, in buffer with galactose oxidase, and then to label treated scraped cells with NaB(3)H(4). After 20 hr interaction between chrysotile asbestos and Syrian hamster cell cultures, an alteration in surface labeling of glycolipids and glycoproteins was observed. Tritiated disialogangliosides (G(Dla)) and the higher molecular weight labeled glycoproteins were significantly reduced by asbestos treatment. Similar chrysotile asbestos-treated cultures were grown in monolayers in MEM (Eagles) with 10% fetal bovine serum for 2, 24, 48, and 72 hr and then surface-labeled with galactose oxidase-. NaB(3)H(4) in phosphate buffer. Little or no difference was observed between surface-labeled lipid or protein distribution in untreated cells and those treated with asbestos for 2 hr. Asbestos-induced polar and neutral glycolipid pattern changes were observed at 24, 48, and 72 hr. Disialo- and trisialogangliosides (the more complex gangliosides) were decreased 85%, whereas globoside GL-4 was decreased by 60% at 72 hr. An overall decrease of labeled glycoproteins was observed at 24-48 hr. By 72 hr there was a complete loss of labeled protein bands with 80,000 dalton molecular mass. Since the changes in glycoproteins and glycolipids occur only after extended exposure of the cells to asbestos, the present studies support the concept that a metabolic rather than immediate masking effect is involved. Comparisons of treatment of Syrian hamster embryo cells with various asbestos fibers for 48 hr in the order of decreasing reduction in complex gangliosides were crocidolite>chrysotile (intermediate)>amosite. Effects of the above fibers on high molecular weight glycoproteins labeling followed the same order. The labeling pattern is reminiscent of the increased simplification of glycolipids and glycoproteins found in transformed cells. In the case of asbestos which appears to have no independent mutagenic capability, it is more likely that the membrane changes induced by asbestos serve to allow other mutagens to pass into the cell so as to act on the nuclear structure.
Animals Asbestos