RNAi screening of the kinome identifies modulators of cisplatin response in ovarian cancer cells
RNAi screening of the kinome identifies modulators of cisplatin response in ovarian cancer cells
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DOI：
10.1016/j.ygyno.2010.05.006
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发表时间：
2010-09-01
影响因子：
4.7
通讯作者：
Azorsa, David O.
中科院分区：
文献类型：
--
作者：
Arora, Shilpi;Bisanz, Kristen M.;Azorsa, David O.
Objective.卵巢癌是女性妇科恶性肿瘤中预后较差的一种。它约占女性所有恶性肿瘤的3%，占所有女性癌症相关死亡的5%。标准的治疗是肿瘤细胞减灭术，然后进行辅助化疗，并在复发时用铂类化疗进行再治疗。为了提高顺铂在卵巢癌细胞中的反应，我们利用高通量RNAi筛选来鉴定激酶调节剂。利用靶向572个激酶的siRNA文库进行高通量RNAi筛选，以鉴定卵巢癌细胞株SKOV 3中顺铂反应的增强剂。RNAi筛选鉴定了至少55种siRNA，其增强顺铂在SKOV3细胞中的生长抑制作用。ATR和CHK1的抑制导致顺铂反应的最大调节。顺铂在siRNA存在下的药物剂量反应验证了这些靶基因的作用。为了表明siRNA数据可以成功地转化为潜在的治疗策略，CHK1进一步用小分子抑制剂PD 407824与顺铂组合靶向。结果显示，CHK1抑制剂PD 407824处理SKOV3和OVCAR3细胞后，卵巢癌细胞对顺铂敏感。我们的数据提供了激酶靶点，可用于为卵巢癌患者设计更好的治疗方法。我们还证明了高通量RNAi筛选的有效性，作为一种工具，用于确定敏感的目标，已知的和建立的化疗药物。(C)2010年爱思唯尔公司All rights reserved.
Objective. Ovarian cancer retains a poor prognosis among the female gynaecological malignancies. It constitutes about 3% of all malignancies in women and accounts for 5% of all female cancer related deaths. A standard treatment is cytoreductive surgery followed by adjuvant chemotherapy, and re-treatment with platinum based chemotherapy at the time of relapse. In order to improve cisplatin response in ovarian cancer cells, we utilized a high-throughput RNAi screening to identify kinase modulators.Methods. A high-throughput RNAi screen was performed using a siRNA library targeting 572 kinases to identify potentiators of cisplatin response in the ovarian cancer cell line SKOV3.Results. RNAi screening identified at least 55 siRNAs that potentiated the growth inhibitory effects of cisplatin in SKOV3 cells. Inhibition of ATR and CHK1 resulted in the greatest modulation of cisplatin response. Drug dose response of cisplatin in the presence of siRNA validated the effects of these target genes. To show that the siRNA data could be successfully translated into potential therapeutic strategies, CHK1 was further targeted with small molecule inhibitor PD 407824 in combination with cisplatin. Results showed that treatment of SKOV3 and OVCAR3 cells with CHK1 inhibitor PD 407824 led to sensitization of ovarian cancer cells to cisplatin.Conclusions. Our data provides kinase targets that could be exploited to design better therapeutics for ovarian cancer patients. We also demonstrate the effectiveness of high-throughput RNAi screening as a tool for identifying sensitizing targets to known and established chemotherapeutic agents. (C) 2010 Elsevier Inc. All rights reserved.