Novel Approach to Identify Genes that Modulate Response of Human Ovarian Cancer Cells to Chemotherapeutic Agents Using High-Throughput RNA Interference

摘要

The application of HT-RNAi represents an innovative functional genomic strategy to rapidly identify important genes involved in the response of cancer cells to the chemotherapeutic agents. We developed a novel HT-RNAi screening assay using a kinase siRNA library to identify genes that modulate the response of ovarian cancer cells to cisplatin and paclitaxel. HT-RNAi assays were conducted using the cell line SKOV-3 to identify genes that sensitize cells to low dose cisplatin. Similar screening assays were done using low dose paclitaxel. Analysis of the screening data resulted in lists of sensitizing hits. Validation of the genes identified as sensitizers was initiated and we confirmed the kinases CHK1 and ATR as sensitizing targets to cisplatin. Validation of the genes that sensitize to paclitaxel is currently underway. Further confirmation of gene silencing by the functional siRNA also showed that CHK1 is being significantly silenced by siRNA targeting CHK1 and further silencing confirmation is in progress. Identification and confirmation of kinases involved in modulating the response to cisplatin and paclitaxel will provide the basis for the development of more effective therapeutic strategies in the treatment of ovarian tumors.