The DNA damage mark pH2AX differentiates the cytotoxic effects of small molecule HDAC inhibitors in ovarian cancer cells
The DNA damage mark pH2AX differentiates the cytotoxic effects of small molecule HDAC inhibitors in ovarian cancer cells
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DOI：
10.4161/cbt.12.6.15956
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发表时间：
2011-09-15
影响因子：
3.6
通讯作者：
Khabele, Dineo
中科院分区：
文献类型：
--
作者：
Wilson, Andrew J.;Holson, Edward;Khabele, Dineo
高级别上皮性卵巢癌对DNA损伤铂类化疗相对敏感，这表明卵巢肿瘤对DNA损伤反应途径的依赖性可用于治疗目的。我们的目标是确定DNA损伤标记γ-H2 AX磷酸化（pH 2AX）是否可用于鉴定用于卵巢癌治疗的合适的细胞毒性组蛋白脱乙酰酶抑制剂（HDACi）。在7种卵巢癌细胞系中测试了19种化学上不同的HDACi化合物。进行荧光、生物化学和基于细胞的测定以评估通过pH 2AX诱导的DNA损伤并测量细胞活力和凋亡。计算了pH 2AX与细胞活力和凋亡的细胞效应之间的关系。将选择的HDACi与顺铂和其他DNA损伤剂组合测试以确定HDACi是否改善DNA损伤剂的作用。HDACi化合物诱导不同水平的pH 2AX表达。HDACi处理的卵巢癌细胞中高水平的pH 2AX与细胞活力降低和细胞凋亡增加密切相关。因此，选择了基于酮的HDACi，并发现其增强顺铂的作用，即使在对DNA损伤药物具有极端抗性的卵巢癌细胞中也是如此。总之，基于荧光的pH 2AX测定可用于确定体外对HDACi的细胞反应，并且可能是鉴定用于治疗卵巢癌的潜在更有效的HDACi的有用工具。此外，这些结果支持将酮衍生的HDACi化合物纳入未来的开发。
High grade epithelial ovarian cancers are relatively sensitive to DNA damaging platinum-based chemotherapy, suggesting that the dependencies of ovarian tumors on DNA damage response pathways can be harnessed for therapeutic purposes. Our goal was to determine if the DNA damage mark gamma- H2AX phosphorylation (pH2AX) could be used to identify suitable cytotoxic histone deacetylase inhibitors (HDACi) for ovarian cancer treatment. Nineteen chemically diverse HDACi compounds were tested in 7 ovarian cancer cell lines. Fluorescent, biochemical and cell-based assays were performed to assess DNA damage by induction of pH2AX and to measure cell viability and apoptosis. The relationships between pH2AX and the cellular effects of cell viability and apoptosis were calculated. Selected HDACi were tested in combination with cisplatin and other DNA damaging agents to determine if the HDACi improved upon the effects of the DNA damaging agents. The HDACi compounds induced differing levels of pH2AX expression. High levels of pH2AX in HDACi-treated ovarian cancer cells were tightly associated with decreased cell viability and increased apoptosis. Consequently, a ketone-based HDACi was chosen and found to enhance the effects of cisplatin, even in ovarian cancer cells with extreme resistance to DNA damaging drugs. In conclusion, a fluorescent-based assay for pH2AX can be used to determine cellular responses to HDACi in vitro and may be a useful tool to identify potentially more effective HDACi for the treatment of ovarian cancer. In addition, these results lend support to the inclusion of ketone-derived HDACi compounds for future development.