The effect of nitrogen mustard (2-chloro-N-2-chloroethyl-N-methylethanamine), Trenimon (2,3,5-trisethyleneiminobenzoquinone-1,4), chlorambucil (4-[p-(bis[2-chloroethyl]amino)-phenyl]butyric acid) and phosphamide mustard (N,N-bis(2-chloroethyl)-diamidophosphoric acid) on Na+/K+-ATPase, membrane fluidity and cell multiplication was studied. With the exception of chlorambucil which does not affect Na+/K+-ATPase all concentrations of the other alkylating agents which inhibit cell multiplication of Ehrlich ascites tumor cells depress the activity of the Na+/K+-ATPase. All alkylating agents--including chlorambucil--caused an increase in the apparent degree of fluorescence polarization after labelling of the plasma membrane with 1,6-diphenyl-1,3,5-hexatriene (DPH). This effect is interpreted as a decrease in membrane fluidity caused by the alkylating drugs. The decrease in membrane fluidity is due to a direct interaction of the alkylating agent with the plasma membrane and is expressed at all concentrations of the drug which inhibit cell proliferation. No effect on membrane fluidity is observed after treatment of cells resistant to nitrogen mustard. The biological consequence of a decrease in membrane fluidity was investigated by growing Friend erythroleukemia cells in the presence of 10 mM cholesterol hemisuccinate. This procedure raises the microviscosity of the plasma membrane and depresses cell proliferation.
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