Differences in membrane order between C3H 10 T1/2 cells and their transformed counterparts as measured by EPR

Membrane order of mouse embryo fibroblasts and their ionizing radiation and chemically transformed counterparts was investigated using EPR spectroscopy after labeling the membrane of the cells with the fatty acid spin label, 5-nitroxy stearic acid. The EPR spectra were recorded at temperatures ranging from 18 to 38 degrees C for both control and transformed cells. The distance between the outer hyperfine splitting (2T' parallel), which is used as indicator of membrane order, varies in these two cell types. Below 28 degrees C. 2T' parallel is higher in transformed fibroblasts than in normal cells, whereas above this temperature membrane order is the same. Lipid analysis as carried out by the measurement of the cholesterol/membrane proteins and sphingomyelin/lecithin ratios, showed no difference in the amounts of the main membrane rigidifiers. These findings suggest that cell transformation of mouse fibroblasts induced by radiation or chemicals may produce alterations in the cell membrane, as evidenced by variations in its order at low temperature. These measured differences are presumably not attributable to its fatty acids composition but to its glycoproteins content, since changes in membrane rigidifiers were not observed between normal and transformed cells.