Effects of N-methylformamide on the growth, cell cycle, and glutathione status of murine TLX5 lymphoma cells.
The growth of the murine TLX5 lymphoma is inhibited in vivo by administration of N-methylformamide (Gescher, A., et al., Br. J. Cancer, 45: 843-850, 1982). Continuous incubation of TLX5 murine lymphoma cells in vitro with N-methylformamide for 72 h, at concentrations of between 43 and 170 mM (0.25 and 1% v/v), brought about a concentration-dependent decrease in growth rate (50% inhibitory concentration = 68 mM) and viability. Cell replication was decreased by 37% after 48 h exposure to 106 mM N-methylformamide, while viability was maintained at 82%. Analysis of the distribution of these cells in the cell cycle by flow cytofluorimetry showed a 23% increase in the proportion of G1 cells and a fall in the proportion of cells in the S and G2/M phases. As the drug concentration and time of exposure to N-methylformamide were increased, with an associated reduction in cell replication and viability, the proportion of G1 cells rose. When TLX5 cells were washed free of N-methylformamide after an exposure to 106 mM for 48 h and cultured in drug-free medium, the cells returned to exponential growth and to a normal cell cycle distribution. Clonogenic assays showed that the recovery of proliferation, after removal of the drug, was due to that of all those cells which, in a parallel experiment, excluded the dye trypan blue. It is concluded that the cessation of replication and the accumulation of cells in G1 of the cell cycle, after treatment with N-methylformamide, are probably not events representative of terminal differentiation but rather of cytostasis, which was accompanied by rapid cell death. Coincident with the reduction of TLX5 cell proliferation caused by N-methylformamide and the accumulation of cells in G1, cellular glutathione concentrations fell by 80%. A similar fall was induced by treatment of the cells with D,L-buthionine[S,R]-sulfoximine (5 microM) for 48 h, but this treatment had no effect on cell growth.
Bill
C A
CA
Pharmaceutical Sciences Institute, Aston University, Birmingham, United Kingdom.
Gescher
A
A
Hickman
J A
JA
D000818:Animals; D019328:Buthionine Sulfoximine; D002453:Cell Cycle / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D005559:Formamides / Q000494:pharmacology*; D005978:Glutathione / Q000378:metabolism*; D008223:Lymphoma / Q000378:metabolism / Q000473:pathology*; D008717:Methionine Sulfoximine / Q000031:analogs & derivatives / Q000494:pharmacology; D051379:Mice; D008808:Mice, Inbred CBA; D014407:Tumor Cells, Cultured
D005559:Formamides; D008717:Methionine Sulfoximine; D019328:Buthionine Sulfoximine; D005978:Glutathione; C002950:methylformamide