Comparative analysis of DNA breakage, chromosomal aberrations and apoptosis induced by the anti-herpes purine nucleoside analogues aciclovir, ganciclovir and penciclovir.
Nucleoside analogues have been used in antiviral therapy and suicide cancer gene therapy. Therefore, it is of importance to compare their potential cytotoxic and genotoxic action. Using metabolically competent CHO cells expressing the thymidine kinase gene of herpes simplex virus type 1 (CHO-HSVtk cells) as a model system, the induction of DNA breaks was compared with the induction of structural chromosomal aberrations and apoptosis/necrosis after exposure to the anti-herpes nucleoside analogues aciclovir (ACV), ganciclovir (GCV) and penciclovir (PCV). After continuous treatment of CHO-HSVtk cells with the drugs, LD(10) in a colony-forming assay was 50, 0.5 and 1 microM for ACV, GCV and PCV, respectively, with GCV to be the most potent agent as determined at a given dose level. There was a remarkable difference in the activity of the agents to kill HSVtk expressing and non-expressing cells: the difference in cellular sensitivity of HSVtk(+) versus HSVtk(-) cells at LD(10) level was 7-fold for ACV, 60-fold for GCV and 400-fold for PCV. The drugs were shown to be strong inducers of apoptosis that was analysed as to concentration- and time-dependence; they induced to only very low extent necrosis. The agents were also highly potent in the induction of DNA single-strand breaks (SSBs) and double-strand breaks (DSBs) (as measured by single cell gel electrophoresis (SCGE)) and chromosomal aberrations. Although PCV induced DNA DSBs with a kinetics and frequency similar to that of GCV, it caused mostly condensation defects instead of "typical" structural chromosomal aberrations. For the drugs used, the frequency of apoptotic cells and the induction of abnormal mitoses appear to be related indicating genotoxic effects induced by the agents to be involved in cell killing due to apoptosis.
Tomicic
Maja T
MT
Institute of Toxicology, Division of Applied Toxicology, University of Mainz, Obere Zahlbacher Str. 67, Germany.
Bey
Erato
E
Wutzler
Peter
P
Thust
Rudolf
R
Kaina
Bernd
B
D000212:Acyclovir / Q000031:analogs & derivatives* / Q000633:toxicity*; D000818:Animals; D000998:Antiviral Agents / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects*; D016466:CHO Cells / Q000187:drug effects; D002869:Chromosome Aberrations; D002875:Chromosomes / Q000187:drug effects*; D003114:Colony-Forming Units Assay; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000187:drug effects; D004249:DNA Damage; D004791:Enzyme Inhibitors / Q000633:toxicity; D015774:Ganciclovir / Q000633:toxicity*; D006147:Guanine; D009336:Necrosis; D018139:Simplexvirus / Q000201:enzymology; D013937:Thymidine Kinase / Q000037:antagonists & inhibitors
D000998:Antiviral Agents; D004791:Enzyme Inhibitors; C053539:penciclovir; D006147:Guanine; D004247:DNA; D013937:Thymidine Kinase; D015774:Ganciclovir; D000212:Acyclovir