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Effect of Methyl Methanesulfonate on the Growth of P388 Lymphoma Cells In Vitro and on Their Rate of Progress through the Cell Cycle¹

Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester 20, England

The effect of methyl methanesulfonate on the growth kinetics of the cloned line P388F of the P388 lymphoma in vitro in suspension culture was studied. The proportion of live and dead cells, mitotic index, labeling index, percentage of labeled mitoses, and grain counts were examined in an asynchronous population, during the period of departure from exponential growth. From this data, it was shown that the S (DNA-synthetic) period was the most sensitive phase of the cell cycle. DNA synthesis from labeled thymidine was increased and the duration of the S was prolonged. The rate of passage of cells from S to G₂ was more affected than that from G₁ to S, but both were reduced relative to control rates. Cells in G₂ at the time of treatment were not delayed through the first post-treatment mitosis but were delayed through the subsequent G₁. Cells in G₁ at the time of treatment were delayed in their passage through the subsequent S period. This alkane sulfonate induced periods of increased mitotic activity, each of which was followed by a wave of increased cell death. Recovery to exponential growth at the control rate occurred four days after treatment.

¹ This work was supported by grants from the British Empire Cancer Campaign and The Medical Research Council.

Received 2/14/67. Accepted 5/29/67.