JaLCDOI 10.18926/AMO/32639
フルテキストURL fulltext.pdf
著者 Zhang, Bo| Seki, Shuji| Akiyama, Kosuke| Tsutsui, Ken| Li, Ting| Nagao, Kazutaka|
抄録 <p>DNA damage induced by cis-diamminedichloroplatinum (II) (cisplatin: cis-DDP), an anticancer drug, was studied in vitro by monitoring the drug-induced conformational change of pUC18 plasmid DNA, the sensitivity to some restriction enzymes of the damaged DNA and the sequence-dependent termination of DNA synthesis caused by cisplatin. Closed circular, superhelical pUC18 DNA was treated at 37 degrees C for 16 h with various concentrations of cisplatin. Cisplatin-dose-dependent conformational change due to unwinding of the treated DNA was detected by agarose gel electrophoresis. To analyze the base-specificity of the cisplatin damage, the measurement for sensitivity of cisplatin-treated DNA to various types of restriction enzyme and sequence gel analysis of the treated DNA were conducted. The results suggested that cisplatin attacked preferentially the sequence of GG &#62; AG &#62; GNG in the order. In the present assay condition, the cisplatin/DNA nucleotide ratios required for the DNA damage detection were roughly 0.025 for the conformational analysis, 0.001 or more for the restriction enzyme analysis, and less than 0.001 for the sequence gel analysis. By using the present method, it was demonstrated that the cisplatin-mediated DNA damage was inhibited by NaCl, KCl, CaCl2 or MgCl2 at their nearly physiological concentrations, and by reducing agents such as thiourea and 2-mercaptoethanol in the reaction mixture.</p>
キーワード DNA damage cisplatin gel electrophoresis sequence gel analysis
Amo Type Article
発行日 1992-12
出版物タイトル Acta Medica Okayama
46巻
6号
出版者 Okayama University Medical School
開始ページ 427
終了ページ 434
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
論文のバージョン publisher
査読 有り
PubMed ID 1336637
Web of Sience KeyUT A1992KE49600004
JaLCDOI 10.18926/AMO/32188
フルテキストURL fulltext.pdf
著者 Zhang, Bo| Watanabe, Sekiko| Akiyama, Kosuke| Li, Ting| Fukushima, Keisuke| Tsutsui, Ken| Seki, Shuji|
抄録 <p>DNA repair synthesis induced in permeable mouse ascites sarcoma cells by peplomycin, an antitumor antibiotic, was studied. Mouse ascites sarcoma (SR-C3H/He) cells were permeabilized with a low concentration of Triton X-100 in an isotonic condition. Permeable cells were treated with an appropriate concentration of peplomycin to introduce single-strand breaks in permeable cell DNA. DNA repair synthesis in peplomycin-treated permeable cells was measured by incubating the cells with four deoxynucleoside triphosphates in an appropriate buffer system. The DNA repair synthesis was enhanced by ATP and NaCl at near physiological concentrations. More than 90% of DNA synthesis in the present system depended on the peplomycin-treatment. The repair nature of the DNA synthesis was confirmed by a BrdUMP density shift technique. The repair patches were largely completed and ligated in the presence of ATP. Analyses using selective inhibitors for DNA polymerases showed that both DNA polymerase Beta and aphidicolin-sensitive DNA polymerases (DNA polymerase alpha and/or delta) were involved in the repair DNA synthesis.&#60;/P&#62;</p>
キーワード DNA repair peplomycin DNA polymerases permeable mouse cells
Amo Type Article
発行日 1991-04
出版物タイトル Acta Medica Okayama
45巻
2号
出版者 Okayama University Medical School
開始ページ 89
終了ページ 94
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
論文のバージョン publisher
査読 有り
PubMed ID 1714230
Web of Sience KeyUT A1991FL60800004