Adriamycin, 10(-8) - 10(-5) M, inhibited transplasma membrane electron transport of uninduced HL-60 cells susceptible to adriamycin and not in uninduced HL-60 cells resistant to adriamycin as measured by reduction of external ascorbate free radical. Electron flow across the plasma membrane was measured with the intact living cells by means of a simple assay procedure whereby the transported electrons were captured by ascorbate free radical to slow the rate of chemical oxidation of ascorbate. The response to adriamycin was rapid with maximum inhibition in less than 1 min. Preincubation was not required and the inhibition presumably was not mediated through effects on DNA replication or transcription. Except at the highest concentration tested of 10 microM, both transplasma membrane electron transport and growth were unaffected by adriamycin with a line of HeLa cells resistant to the drug. The findings provide evidence, using a physiological acceptor, ascorbate free radical, for a direct inhibition of transmembrane electron transport of HL-60 cells by adriamycin that correlates closely with adriamycin inhibition of cell growth. The lack of response with resistant cells suggests an alternative mechanism for adriamycin resistance not necessarily based on transport control.