Calcium‐dependent stimulation of BALB/c 3T3 mouse cell DNA synthesis by a tumor‐promoting phorbol ester (PMA)

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Abstract
During the first six passages after their arrival in this laboratory, BALB/c 3T3 mouse cells did not proliferate in serum containing‐medium having an ionic calcium concentration of 0.05 mM or less, but by the ninth passage they had become able to multiply in the presence of these lower calcium levels. In low calcium (e.g., 0.02 mM) medium, passage 1–6, cells in sparse cultures were blocked at the Gl/S boundary of their cycle. These blocked cells could be induced to start making DNA within only one hour either by returning the ionic calcium level to a normal range of values (1.25 mM), or by adding 0.05 μg/ml of PMA (12‐O‐tetradecanoyl‐phorbol‐13‐acetate). PMA probably acted by sensitizing the blocked cells to calcium rather than replacing the ion, because it was ineffective in ionic calcium‐free medium. Finally, PMA did not by itself induce proliferation of cells (regardless of the number of passages) which had been proliferatively inactivated by density‐dependent factors in confluent cultures. However, PMA did promote DNA synthesis by these cells during their brief transition to the “cycling” state caused by exposure to fresh serum‐containing medium.

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