Department of Microbiology and Immunology, Georgetown University, Washington, District of Columbia, USA.
Cancer Res. 2012 Nov 15;72(22):5945-55. doi: 10.1158/0008-5472.CAN-12-1400. Epub 2012 Aug 31.
Resistance to platinum-based therapies arises by multiple mechanisms, including by alterations to cell-cycle kinases that mediate G(2)-M phase arrest. In this study, we conducted parallel high-throughput screens for microRNAs (miRNA) that could restore sensitivity to cisplatin-resistant cells, and we screened for kinases targeted by miRNAs that mediated cisplatin resistance. Overexpression of the cell-cycle kinases WEE1 and CHK1 occurred commonly in cisplatin-resistant cells. miRNAs in the miR-15/16/195/424/497 family were found to sensitize cisplatin-resistant cells to apoptosis by targeting WEE1 and CHK1. Loss-of-function and gain-of-function studies showed that miR-15 family members controlled the expression of WEE1 and CHK1. Supporting these results, we found that in the presence of cisplatin altering expression of miR-16 or related genes altered cell cycle distribution. Our findings reveal critical regulation of miRNAs and their cell-cycle-associated kinase targets in mediating resistance to cisplatin.
铂类治疗耐药性是通过多种机制产生的，包括细胞周期激酶的改变，这些激酶介导 G2-M 期阻滞。在这项研究中，我们进行了平行的高通量筛选，以寻找能够恢复顺铂耐药细胞敏感性的 microRNAs（miRNA），并筛选由 miRNA 靶向的介导顺铂耐药的激酶。细胞周期激酶 WEE1 和 CHK1 的过表达在顺铂耐药细胞中普遍发生。发现 miR-15/16/195/424/497 家族中的 miRNA 通过靶向 WEE1 和 CHK1 使顺铂耐药细胞对细胞凋亡敏感。功能丧失和功能获得研究表明，miR-15 家族成员控制 WEE1 和 CHK1 的表达。支持这些结果，我们发现，在顺铂存在的情况下，miR-16 或相关基因的表达改变会改变细胞周期分布。我们的研究结果揭示了 miRNA 及其与细胞周期相关的激酶靶标在介导顺铂耐药性方面的关键调控作用。
