title: The cytotoxic and anti-proliferative effects of 3-hydrogenkwadaphnin in K562 and jurkat cells is reduced by guanosine.
abstract: 3-hydrogenwadaphnin (3-HK) is a new daphnane-type diterpene ester isolated from Dendrostellera lessertii with strong anti-tumoral activity in animal models and in cultures. Here, prolonged effects of this new agent on proliferation and viability of several different cancerous cell lines were evaluated. Using [(3)H]thymidine incorporation, it was found that the drug inhibited cell proliferation and induced G1/S cell cycle arrest in leukemic cells 24 h after a single dose treatment. The cell viability of Jurkat cells was also decreased by almost 10 %, 31 % and 40 % after a single dose treatment (7.5 nM) at 24, 48 and 72 h, respectively. The drug-treated cells were stained with acridine orange/ ethidium bromide to document the chromatin condensation and DNA fragmentation. These observations were further confirmed by detection of DNA laddering pattern in the agarose gel electrophoresis of the extracted DNA from the treated cells. Treatment of K562 cells with the drug at 7.5, 15 and 30 nM caused apoptosis in 25 %, 45 % and 65 % of the cells, respectively. Exogenous addition of 25-50 microM guanosine and/or deoxyguanosine to the cell culture of the drug-treated cells restored DNA synthesis, released cell arrest at G1/S checkpoint and decreased the apoptotic cell death caused by the drug. These observations were not made using adenosine. However, the drug effects on K562 cells were potentiated by hypoxanthine. Based on these observations, perturbation of GTP metabolism is considered as one of the main reasons for apoptotic cell death by 3-HK.
authors.affiliation: Institute of Biochemistry and Biophysics, P. O. Box. 13145-1384, University of Tehran, Tehran, Iran.
authors.forename: Mohammad Amin
authors.initials: MA
authors.lastname: Moosavi
authors.forename: Razieh
authors.initials: R
authors.lastname: Yazdanparast
authors.forename: Mohammad Hasan
authors.initials: MH
authors.lastname: Sanati
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002843:Chromatin / Q000378:metabolism; D003849:Deoxyguanosine / Q000494:pharmacology; D004224:Diterpenes / Q000494:pharmacology*; D016193:G1 Phase / Q000187:drug effects*; D006151:Guanosine / Q000494:pharmacology*; D006801:Humans; D019169:Jurkat Cells; D020014:K562 Cells; D008519:Medicine, Traditional; D010936:Plant Extracts / Q000494:pharmacology*; D029645:Thymelaeaceae / Q000737:chemistry; D013936:Thymidine / Q000378:metabolism
chemical_list: C505417:3-hydrogenkwadaphnin; D000970:Antineoplastic Agents; D002843:Chromatin; D004224:Diterpenes; D010936:Plant Extracts; D006151:Guanosine; D003849:Deoxyguanosine; D013936:Thymidine