Pharmaceutical Genomics Division, The Translational Genomics Research Institute, Scottsdale, AZ 85259, USA.
Gynecol Oncol. 2010 Sep;118(3):220-7. doi: 10.1016/j.ygyno.2010.05.006.
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.
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.
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.
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.
在女性妇科恶性肿瘤中，卵巢癌的预后仍然较差。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%。标准治疗是细胞减灭术，继以辅助化疗，并在复发时用铂类为基础的化疗重新治疗。为了提高卵巢癌细胞对顺铂的反应，我们利用高通量 RNAi 筛选来鉴定激酶调节剂。
使用针对 572 种激酶的 siRNA 文库对卵巢癌细胞系 SKOV3 进行高通量 RNAi 筛选，以鉴定顺铂反应的增强剂。
RNAi 筛选鉴定出至少 55 种 siRNA 增强了 SKOV3 细胞中顺铂的生长抑制作用。ATR 和 CHK1 的抑制导致顺铂反应的最大调节。siRNA 存在时顺铂的药物剂量反应验证了这些靶基因的作用。为了表明 siRNA 数据可以成功转化为潜在的治疗策略，我们用小分子抑制剂 PD 407824 进一步靶向 CHK1，并与顺铂联合使用。结果表明，CHK1 抑制剂 PD 407824 处理 SKOV3 和 OVCAR3 细胞导致卵巢癌细胞对顺铂的敏感性增加。
我们的数据提供了可用于设计针对卵巢癌患者更好治疗方法的激酶靶标。我们还证明了高通量 RNAi 筛选作为鉴定已知和既定化疗药物增敏靶标的有效工具。
