Cell cycle traverse in AHH-1 tk +/- human lymphoblastoid cells exposed to the chromosomal mutagen, m-amsa.
AHH-1 tk +/- cells were exposed to the chemotherapeutic agent, m-amsa, both in complete medium and in medium without serum, subcultured in complete medium, and the effect on the traverse of the cell cycle determined by flow cytometric analysis of bromodeoxyuridine (BrdUrd)-labeled DNA. After exposure to m-amsa (day 0), the percentage of S-phase cells increased significantly (P < 0.0017) with increasing concentration. Cells also accumulated in G2/M as evidenced by the significant (P < 0.0026), concentration-dependent increase in the percentage of cells detected within this phase. Serum deprivation during exposure resulted in significantly (P = 0.024) more cells in S-phase than in cultures exposed to m-amsa in complete medium. After three days in culture, a significant (P = 0.0001) accumulation of cells in G2/M was present; the percentage of cells in G2/M did not differ significantly (P = 0.148) in cultures exposed to m-amsa in complete medium or in serum-free medium. However, a significant (P < 0.001) loss of S-phase cells was found in cultures exposed without serum. At day 7, no significant concentration effects were detected (GO/G1, P = 0.6026; S-phase, P = 0.9773; G2/M, P = 0.8401). These results demonstrate that exposure to m-amsa perturbs the traverse of the cell cycle, initially by inhibiting the completion of S-phase and followed by an accumulation of cells in G2/M. In addition, exposure to m-amsa under conditions of serum deprivation results in an increased percentage of cells in the initial S-phase after exposure, the loss of S-phase cells from the culture after three days, and the appearance of subdiploid peak, consistent with cells undergoing apoptosis.
Morris
S M
SM
Division of Genetic Toxicity, Food and Drug Administration, National Center for Toxicological Research, Jefferson, Arkansas, USA.
McGarrity
L J
LJ
Domon
O E
OE
Chen
J J
JJ
Casciano
D A
DA
D000677:Amsacrine / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects*; D001402:B-Lymphocytes / Q000187:drug effects*; D002453:Cell Cycle / Q000187:drug effects*; D002461:Cell Line, Transformed; D002877:Chromosomes, Human / Q000187:drug effects; D016895:Culture Media, Serum-Free; D004249:DNA Damage; D004791:Enzyme Inhibitors / Q000633:toxicity*; D005434:Flow Cytometry; D016195:G2 Phase / Q000187:drug effects; D006801:Humans; D007364:Intercalating Agents / Q000633:toxicity*; D009153:Mutagens / Q000633:toxicity*; D013937:Thymidine Kinase / Q000235:genetics; D059005:Topoisomerase II Inhibitors
D016895:Culture Media, Serum-Free; D004791:Enzyme Inhibitors; D007364:Intercalating Agents; D009153:Mutagens; D059005:Topoisomerase II Inhibitors; D000677:Amsacrine; D013937:Thymidine Kinase