R-Ketorolac Targets Cdc42 and Rac1 and Alters Ovarian Cancer Cell Behaviors Critical for Invasion and Metastasis.
R-Ketorolac Targets Cdc42 and Rac1 and Alters Ovarian Cancer Cell Behaviors Critical for Invasion and Metastasis.
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DOI：
10.1158/1535-7163.mct-15-0419
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发表时间：
2015-10
影响因子：
5.7
通讯作者：
Wandinger-Ness A
中科院分区：
文献类型：
--
作者：
Guo Y;Kenney SR;Muller CY;Adams S;Rutledge T;Romero E;Murray-Krezan C;Prekeris R;Sklar LA;Hudson LG;Wandinger-Ness A
Cdc 42（细胞分裂控制蛋白42）和Rac 1（Ras相关的C3肉毒杆菌毒素底物1）是卵巢癌中有吸引力的治疗靶点，这是基于肿瘤细胞迁移、粘附和侵袭的重要性。尽管有预期的益处，但靶向GTP酶尚未转化为临床实践。我们以前建立了Cdc 42和组成型活性Rac 1b在原发性卵巢肿瘤组织中过表达。通过高通量筛选和计算形状同源性方法，我们将R-酮咯酸鉴定为Cdc 42和Rac 1抑制剂;与S-酮咯酸的抗炎、环氧合酶抑制活性不同。在本研究中，我们建立了R-酮咯酸作为Cdc 42和Rac 1的变构抑制剂。基于细胞的测定验证了R-酮咯酸对Cdc 42和Rac 1的活性。对永生化人卵巢腺癌细胞（SKOV 3 ip）和原代患者源性卵巢癌细胞的研究表明，R-酮咯酸是生长因子或血清依赖性Cdc 42和Rac 1激活的强效抑制剂，其效力和细胞功效与Cdc 42（CID 2950007/ML 141）和Rac 1（NSC 23766）的小分子抑制剂相似。此外，R-酮咯酸对GT3的抑制作用使下游p21激活激酶（PAK 1/PAK 2）效应物激活降低> 80%。使用SKOV 3 ip和原代患者来源的卵巢癌细胞进行的细胞行为的多项测定显示，R-酮咯酸显著抑制细胞粘附、迁移和侵袭。总之，我们提供了R-酮咯酸作为Cdc 42和Rac 1直接抑制剂的证据，能够调节下游GTPases依赖性生理反应，这对肿瘤转移至关重要。我们的研究结果表明，FDA批准的药物外消旋酮咯酸可选择性抑制Cdc 42和Rac 1 GTP酶，可用于人体。
Cdc42 (cell division control protein 42) and Rac1 (Ras-related C3 botulinum toxin substrate 1) are attractive therapeutic targets in ovarian cancer based on established importance in tumor cell migration, adhesion and invasion. Despite a predicted benefit, targeting GTPases has not yet been translated to clinical practice. We previously established that Cdc42 and constitutively active Rac1b are overexpressed in primary ovarian tumor tissues. Through high throughput screening and computational shape homology approaches we identified R-ketorolac as a Cdc42 and Rac1 inhibitor; distinct from the anti-inflammatory, cyclooxygenase inhibitory activity of S-ketorolac. In the present study, we establish R-ketorolac as an allosteric inhibitor of Cdc42 and Rac1. Cell-based assays validate R-ketorolac activity against Cdc42 and Rac1. Studies on immortalized human ovarian adenocarcinoma cells (SKOV3ip), and primary, patient-derived ovarian cancer cells show R-ketorolac is a robust inhibitor of growth factor or serum dependent Cdc42 and Rac1 activation with a potency and cellular efficacy similar to small molecule inhibitors of Cdc42 (CID2950007/ML141) and Rac1 (NSC23766). Furthermore, GTPase inhibition by R-ketorolac reduces downstream p21-activated kinases (PAK1/PAK2) effector activation by >80%. Multiple assays of cell behavior using SKOV3ip and primary patient-derived ovarian cancer cells show that R-ketorolac significantly inhibits cell adhesion, migration and invasion. In sum, we provide evidence for R-ketorolac as direct inhibitor of Cdc42 and Rac1 that is capable of modulating downstream GTPase-dependent, physiological responses, which are critical to tumor metastasis. Our findings demonstrate the selective inhibition of Cdc42 and Rac1 GTPases by an FDA approved drug-racemic ketorolac that can be used in humans.