title: Analysis of the antitumor activity of gemcitabine and carboplatin against ovarian clear-cell carcinoma using the DNA damage marker γH2AX.
abstract: BACKGROUND
  Differences in the incidence and type of DNA damage induced by antitumor agents for ovarian clear-cell carcinoma (CCC) were determined in two CCC cell lines, using γH2AX.
  
  
  METHODS
  The antitumor activity of gemcitabine (GEM) and carboplatin (CBDCA) were examined using cultured cell lines of CCC (OVISE and RMG-I). Each cell line was treated with GEM and CBDCA, the cells were collected, fixed, and then reacted with anti-γH2AX antibody. γH2AX and nuclear DNA were then simultaneously detected by flow cytometry using fluorescein isothiocyanate and propidium iodide, respectively, to determine the amounts of γH2AX formed in each cell-cycle phase.
  
  
  RESULTS
  After administration of GEM, both cell lines showed DNA damage and cell-cycle arrest in the S and G2/M phases, and increased apoptosis. Similarly, with CBDCA, OVISE showed S- and G2/M-phase arrest, while RMG-I showed G2/M-phase arrest.
  
  
  CONCLUSIONS
  The mechanism of action of GEM and CBDCA in CCC cell lines was elucidated using γH2AX as a DNA damage marker. Our findings suggested that concomitant use of GEM plus CBDCA may be effective in the treatment of CCC.
authors.affiliation: Department of Obstetrics and Gynecology.
authors.forename: Eriko
authors.initials: E
authors.lastname: Takatori
authors.forename: Tadahiro
authors.initials: T
authors.lastname: Shoji
authors.forename: Takashi
authors.initials: T
authors.lastname: Sawai
authors.forename: Akira
authors.initials: A
authors.lastname: Kurose
authors.forename: Toru
authors.initials: T
authors.lastname: Sugiyama
keywords: DNA damage; apoptosis; carboplatin; clear-cell carcinoma; gemcitabine; ovarian cancer; γH2AX
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