Detection of an early surface change during oncogenic transformation.

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Abstract
Fluorescamine, which can label surface components of cells grown as monolayers in culture, was used to probe alterations in chicken embryo fibroblasts infected with a temperature-sensitive mutant of Rous sarcoma virus, Prague A, LA24. The fluorescence of bound fluorescamine on cells at the permissive temperature (35.degree. C) was about 1/3 that of cells cultured at the nonpermissive temperature (41.degree. C). During development of the transformed phenotype, i.e., after transfer of the cells from 41.degree. C to 35.degree. C, the decrease in surface fluorescence was observed to be an early event, occurring within the first 4 to 8 h after temperature shift. This alteration took place on a time scale similar to that of changes in 2-deoxyglucose transport and an increased rate of DNA synthesis, but before any major morphological changes. The change was related to cell transformation rather than to growth differences of the cells at the 2 temperatures. Fluorescamine was not monitoring the loss of LETS glycoprotein from the surface or the loss of any other surface components that could be detected by lactoperoxidase-catalyzed iodination of surface proteins. When fluorescamine-labeled components were resolved by polyacrylamide gel electrophoresis, significant differences were seen between components from cells cultured at 35.degree. C compared with those from cells cultured at 41.degree. C. Based on these results, possible mechanisms accounting for the fluorescence differences are suggested.

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