DeMoe Joanna H, Santaguida Stefano, Daum John R, Musacchio Andrea, Gorbsky Gary J
Cell Cycle and Cancer Biology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104, USA.
Cancer Res. 2009 Feb 15;69(4):1509-16. doi: 10.1158/0008-5472.CAN-08-3133. Epub 2009 Feb 3.
In mitosis, the kinetochores of chromosomes that lack full microtubule attachments and/or mechanical tension activate a signaling pathway called the mitotic spindle checkpoint that blocks progression into anaphase and prevents premature segregation of the chromatids until chromosomes become aligned at the metaphase plate. The spindle checkpoint is responsible for arresting cells in mitosis in response to chemotherapeutic spindle poisons such as paclitaxel or vinblastine. Some cancer cells show a weakened checkpoint signaling system that may contribute to chromosome instability in tumors. Because complete absence of the spindle checkpoint leads to catastrophic cell division, we reasoned that drugs targeting the checkpoint might provide a therapeutic window in which the checkpoint would be eliminated in cancer cells but sufficiently preserved in normal cells. We developed an assay to identify lead compounds that inhibit the spindle checkpoint. Most cells respond to microtubule drugs by activating the spindle checkpoint and arresting in mitosis with a rounded morphology. Our assay depended on the ability of checkpoint inhibitor compounds to drive mitotic exit and cause cells to flatten onto the substrate in the continuous presence of microtubule drugs. In this study, we characterize one of the compounds, OM137, as an inhibitor of Aurora kinases. We find that this compound is growth inhibitory to cultured cells when applied at high concentration and potentiates the growth inhibitory effects of subnanomolar concentrations of paclitaxel.
在有丝分裂过程中,缺乏完整微管附着和/或机械张力的染色体的动粒会激活一种名为有丝分裂纺锤体检查点的信号通路,该通路会阻止细胞进入后期,并防止染色单体过早分离,直到染色体在中期板上排列整齐。纺锤体检查点负责在细胞对化疗性纺锤体毒素(如紫杉醇或长春碱)作出反应时,使细胞停滞在有丝分裂阶段。一些癌细胞显示出减弱的检查点信号系统,这可能导致肿瘤中的染色体不稳定。由于完全缺失纺锤体检查点会导致灾难性的细胞分裂,我们推测,靶向该检查点的药物可能会提供一个治疗窗口,在这个窗口中,癌细胞中的检查点将被消除,而在正常细胞中则能得到充分保留。我们开发了一种检测方法来鉴定抑制纺锤体检查点的先导化合物。大多数细胞通过激活纺锤体检查点并以圆形形态停滞在有丝分裂中来响应微管药物。我们的检测方法依赖于检查点抑制剂化合物在微管药物持续存在的情况下驱动有丝分裂退出并使细胞扁平贴附在底物上的能力。在这项研究中,我们将其中一种化合物OM137鉴定为极光激酶的抑制剂。我们发现,当高浓度应用该化合物时,它对培养细胞具有生长抑制作用,并能增强亚纳摩尔浓度紫杉醇的生长抑制效果。