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Published on: July 3, 2015
DNA damage detected with gammaH2AX in endometrioid adenocarcinoma cell lines
Maki Ikeda1, Akira Kurose, Eriko Takatori
1Department of Pathology, Iwate Medical University School of Medicine, Morioka, Iwate 020-8505, Japan.
Abstract:
Phosphorylation of histone H2AX (gammaH2AX) is a sensitive marker of DNA damage, particularly induction of DNA double-strand breaks. Using multiparameter cytometry we explored the effects of doxorubicin (DOX), cisplatin (CDDP) and 5-fluorouracil (5-FU) on four types of endometrioid adenocarcinoma cell lines (HEC-1A, HEC-1B, Ishikawa, KLE) correlating the drug-induced increases in phosphorylated H2AX (gammaH2AX) with cell cycle phase, induction of apoptosis and induction of cell senescence, the latter detected by analysis of beta-galactosidase. The study revealed significant differences among the cell lines in the effects of DNA damage vis-a-vis cell cycle phase specificity, induction of apoptosis or senescence following drug treatment. DOX treatment showed little cell cycle specificity in terms of induction of gammaH2AX, and its mechanism, which is similar to another anthracycline DNA topoisomerase II inhibitor mitoxantrone, may involve oxidative DNA damage modulated by other factors. Treatment with CDDP and 5-FU led to phosphorylation of H2AX preferentially in S-phase cells, consistent with the induction of replication stress. The response of Ishikawa cells expressing wt p53 was different compared to other cell lines. The data suggest that the treatment of endometrioid adenocarcinoma with these drugs may have to be customized to individual patients. The flow cytometric bivariate analysis of gammaH2AX and DNA content is a useful technique for better understanding the effects of antitumor agents and may contribute to customized patient treatments.
Insights
DNA damage marker phosphorylated H2AX (gammaH2AX) response varies among endometrioid adenocarcinoma cell lines treated with doxorubicin, cisplatin, and 5-fluorouracil, suggesting personalized cancer therapy is needed.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Phosphorylation of histone H2AX (gammaH2AX) is a key indicator of DNA damage, particularly double-strand breaks.
- Endometrioid adenocarcinoma treatment efficacy can vary significantly due to cellular responses to chemotherapy.
Purpose of the Study:
- To investigate the differential effects of doxorubicin, cisplatin, and 5-fluorouracil on DNA damage response in endometrioid adenocarcinoma cell lines.
- To correlate drug-induced gammaH2AX levels with cell cycle progression, apoptosis, and senescence.
- To explore the potential for customized treatment strategies based on cellular drug sensitivity.
Main Methods:
- Utilized multiparameter flow cytometry to analyze gammaH2AX phosphorylation.
- Assessed cell cycle phase distribution, apoptosis, and beta-galactosidase activity for senescence.
- Compared responses across four endometrioid adenocarcinoma cell lines (HEC-1A, HEC-1B, Ishikawa, KLE).
Main Results:
- Doxorubicin induced gammaH2AX with little cell cycle specificity, suggesting potential oxidative damage mechanisms.
- Cisplatin and 5-fluorouracil treatments preferentially induced gammaH2AX in S-phase cells, indicating replication stress.
- Significant variations in DNA damage response, apoptosis, and senescence were observed among the cell lines.
- Ishikawa cells with wild-type p53 exhibited a distinct response compared to other cell lines.
Conclusions:
- Drug-induced DNA damage and subsequent cellular responses (apoptosis, senescence) differ across endometrioid adenocarcinoma cell lines.
- The cell cycle specificity of DNA damage varies depending on the chemotherapeutic agent.
- Flow cytometric analysis of gammaH2AX is valuable for understanding antitumor agent effects.
- Tailoring chemotherapy for endometrioid adenocarcinoma based on individual patient cell line characteristics is a promising approach for personalized medicine.
