Irreversible G2-M arrest and cytoskeletal reorganization induced by cytotoxic nucleoside analogues.
The mechanisms by which cytotoxic agents perturb the normal cell biology and cell cycle progression of cancer cells were explored using B16F10 cells genetically modified to express the Herpes Simplex Virus-thymidine kinase gene. Culture in the presence of the nucleoside analogue ganciclovir induced a profound morphological change that required entry of treated cells into S phase and was dependent on prenylated proteins such as those of the rho gene family. Cell cycle arrest occurred in late S phase or G2 phase due to the activation of the G2-M DNA damage checkpoint. This checkpoint control operated at the level of inhibition of the activity of Cdc2/cyclin B and occurred by two mechanisms: (a) p53-mediated up-regulation of p21CIP/WAF1 expression and its association with Cdc2/cyclin B; and (b) prevention of the dephosphorylation of tyrosine 15 of Cdc2. These events occurred in vitro and in vivo, and were shown to mediate bystander killing in this model. The mechanism of cell death seemed to be due to the irreversible cell cycle arrest at the G2-M checkpoint, rather than induction of apoptosis. These data link DNA damage checkpoints with cytoskeletal signaling pathways and the core cell cycle machinery and may represent a general mechanism of cytotoxicity of this class of nucleoside analogues.
Halloran
P J
PJ
Department of Experimental immunology and Transplantation, National Cancer Institute-Frederick Cancer Research and Development Center, Maryland 21702, USA.
Fenton
R G
RG
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D019926:Cyclin B / Q000037:antagonists & inhibitors; D003599:Cytoskeleton / Q000187:drug effects*; D004249:DNA Damage; D005260:Female; D016195:G2 Phase / Q000187:drug effects*; D015774:Ganciclovir / Q000494:pharmacology*; D008148:Lovastatin / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D008938:Mitosis / Q000187:drug effects*; D006656:Protamine Kinase / Q000037:antagonists & inhibitors; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D019926:Cyclin B; D008148:Lovastatin; D006656:Protamine Kinase; D015774:Ganciclovir