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Molecular Pharmacology, Vol 18, 274-280, Copyright © 1980 by the American Society for Pharmacology and Experimental Therapeutics

Alterations in the Kinetics of Methotrexate Transport During Growth of L1210 Murine Leukemia Cells in Culture

PAUL L. CHELLO 1, FRANCIS M. SIROTNAK 1, and DIANE M. DORICK 1

1 Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, New York 10021

Measurements of methotrexate transport during different phases of growth of L1210 murine leukemia cells in culture revealed no significant change in the Km for methotrexate influx (3.9-4.7 µM) during growth. However, the Vmax for influx into exponentially growing (midlog) L1210 cells was threefold greater (9.4 nmol/min/g dry wt) than the Vmax for influx into stationary cells (3.1 nmol/min/g dry wt), while the rate of methotrexate efflux from stationary phase cells (K, 0.06 min-1) was twice that measured for midlog cells (K, 0.03 min-1). These reciprocal changes in the kinetic parameters for influx and efflux accounted for a five- to sixfold greater steady-state level for exchangeable methotrexate in midlog cells than in stationary phase cells. These data may help to explain the increased sensitivity of exponentially growing versus stationary phase cells to methotrexate.

Note:
ACKNOWLEDGMENT The authors wish to thank Ms. Ellen Wong for excellent technical assistance.

Submitted on September 24, 1979
Accepted on March 28, 1980




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