.ALPHA.-fetoprotein, DNA, and RNA in human hepatoma cell lines using cytoflowmetry.

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Cells were fixed in cold ethanol solutions for immunohistological staining of AFP by the ABC method. The AFP-positive ratio was determined after fixation in each concentration of EtOH. (of 50, 60, 70, 80, 90, 95 and of 100% EtOH, each for 5 minutes) and also after a stepwise fixation transfering the cells in turn from 50% EtOH. to 100% EtOH. (in 10% EtOH. steps each for 1 minute). The AFP-positive ratio was highest following the stepwise fixation. Also, the contractile tendency decreased at increased EtOH. concentrations. The DNA ploidy pattern was examined for the AFP-positive cells and the AFP-negative cells. In the lag phase and in the logarithmic growth phase, the number of the AFP-positive cells in the S +G2·M phase was higher than that of the AFP-negative cells. In the stationary phase, there was no difference between them. The relationship between AFP content and the phase of the cell cycle was investigated. AFP content increased with the development of the cell cycle, with only a slight decrease in the early, mid S phase. Acridine orange (AO) staining was performed after AFP staining with an indirect immunofluorescent method. As a result, the amount of green fluorescence of DNA was unchanged, while the red fluorescence of RNA was slightly weaker. The AFP content was correlated with the RNA content.
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