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Onco Targets Ther. 2013 Jul 17;6:901-7. doi: 10.2147/OTT.S44021. Print 2013.

Analysis of the antitumor activity of gemcitabine and carboplatin against ovarian clear-cell carcinoma using the DNA damage marker γH2AX.

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  • 1Department of Obstetrics and Gynecology.

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.

MATERIALS AND 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.

CONCLUSION:

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.

KEYWORDS:

DNA damage; apoptosis; carboplatin; clear-cell carcinoma; gemcitabine; ovarian cancer; γH2AX

PMID:
23898228
[PubMed]
PMCID:
PMC3718840
Free PMC Article

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