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Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1990 Jan;81(1):85–90. doi: 10.1111/j.1349-7006.1990.tb02511.x

Methotrexate Cytotoxicity as Related to Irreversible S Phase Arrest in Mouse L1210 Leukemia Cells

Masahito Tsurusawa 1,✉, Makoto Niwa 1, Naoyuki Katano 1, Takeo Fujimoto 1
PMCID: PMC5917961  PMID: 2108951

ABSTRACT

The association between cytotoxicity and cell cycle perturbation caused by methotrexate (MTX) was investigated in mouse L1210 leukemia cells by flow cytometric bromodeoxyuridine/DNA assay. In the range of concentrations of MTX from 10‐7M to 10‐6M, in vitro exposure to the drug for 6 h caused a dose‐dependent suppression of clonal growth of the tumor cells and S phase arrest in the cycle progression, resulting in an accumulation of cells in early S phase, in which they showed no definite increase of DNA content above G1 levels. The surviving fraction of the clonogenic cells corresponded with the fraction of cells which recovered from the S phase arrest in MTX‐free medium. In mice bearing L1210 ascites tumors, a bolus injection of MTX caused the S phase arrest of the tumor cells as shown in suspension cultures, and cytokinetic recovery was observed in parallel with the regrowth of the tumor. These results showed that irreversible S phase arrest is a critical cytokinetic event associated with the cytotoxicity of MTX.

Keywords: Flow cytometry, L1210 leukemia, Methotrexate, S phase arrest

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