By Karl Maramorosch
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5 a Cultures were grown for 72 hours in medium. , 1980). 48 RODY P. COX AND DEBRA GENDUSO DAY above, the ectopic synthesis of glycopeptide hormones in HeLa cells is associated with cessation of cell division. This has been shown for control cultures approaching confluence (Fig. 2) as well as dispersed cell cultures to which sodium butyrate or other inhibitors of DNA synthesis were added (Fig. 2). Since most examples of cell differentiation are associated with blocks in the cell cycle, it seemed important to determine if butyrate focused the cell population.
A. Effects of Sodium Butyrate and Hydroxyurea Proliferation on Cell The kinetics of growth inhibition produced by 1 and 5 m M butyrate and 1 m M hydroxyurea are seen in Fig. 6. Both control (without additions) and cultures with 1 m M butyrate grew exponentially, although the butyrate-treated population had a moderate increase in doubling time, which was experimentally shown to be a prolonged G, phase (Fallon and Cox, 1979). Cell populations grown with 5 m M butyrate and 1 m M hydroxyurea showed a marked inhibition of multiplication, reaching a plateau in 12 hours.
By 14 hours all except 40% of the hydroxyurea-treated cells have already completed S phase and at 24 hours the labeling index is 3%, demonstrating the degree of synchrony attained by this compound. Control HeLa cells maintain a nearly constant proportion of cells in S phase with an increase in labeling index during the longer pulse periods (Table IX). Analysis of cultures grown with 5 m M butyrate shows t h a t following release of inhibition, 60% of cells immediately enter DNA synthesis and the kinetics of labeling and decline are similar to hydroxyurea.