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C. Antczak, C. Radu, E. Kim, H. Djaballah, D. H. Abramson; High-Throughput Identification of Antitumor Agents for Retinoblastoma. Invest. Ophthalmol. Vis. Sci. 2007;48(13):93.
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The purpose of this study is to identify potentially more potent chemotherapeutic agents for retinoblastoma.
We developed an assay allowing the high-throughput screening (HTS) of large chemical libraries. The assay is performed in 1536-well microtiter plates in a total volume of 10 µL. We chose the Y79 cell line as a model of retinoblastoma compatible with the requirements of HTS. The Alamar blue assay we employed relies on the reduction of the resazurin dye by metabolically active cells. The resulting fluorescence is proportional to the number of live cells, which allows to monitor the cytotoxicity of compounds in a high-throughput fashion.
The statistical performance of the assay was asssessed in a control run and demonstrated the robustness of the miniaturized assay (Z’=0.71). The assay was validated in a pilot screen of 2,640 known bioactive chemicals. This study led to the identification of several known cytotoxic agents belonging to different pharmacological classes (including ion pump effectors, topoisomerase inhibitors and microtubule inhibitors) as potent antitumor agents for retinoblastoma cells.
These results demonstrate that the newly developed assay is highly amenable to HTS. The strategy employed in this study should allow the rapid screening of our library of more than 200,000 compounds and potentially identify original antitumor agents for retinoblastoma.
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