Department of Pharmacology and Physiology, The George Washington University Medical Center, Washington, DC 20037, USA.
Carcinogenesis. 2010 May;31(5):785-93. doi: 10.1093/carcin/bgq014. Epub 2010 Jan 20.
Polo-like kinase 1 (Plk1) is a key regulator of mitosis. Aberrant Plk1 activity is found in tumors, but little is known regarding its role in the DNA damage response of normal cells and its potential contribution to the early stages of carcinogenesis. Inappropriate survival signaling after DNA damage may facilitate clonal expansion of genetically compromised cells, and it is known that protein tyrosine phosphatase (PTP) inhibitors activate key survival pathways. In this study, we employed hexavalent chromium [Cr(VI)], a well-documented genotoxicant, to investigate the mechanism by which survival pathway activation could lead to loss of checkpoint control via a mechanism involving Plk1. We recently reported that PTP inhibition enhances clonogenic survival and mutagenesis after Cr(VI) exposure by overriding Cr-induced growth arrest. Here, we report that checkpoint bypass, facilitated by PTP inhibition, was associated with decreased Cdk1 Tyr15 phosphorylation, as well as increased Plk1 activity and nuclear localization. Plk1 was necessary for increased survival after PTP inhibition and Cr(VI) exposure in normal human fibroblasts via enhanced mitotic progression. In addition, pharmacological inhibition of Plk1 abolished the PTP inhibitor-induced bypass of the G(2)/M checkpoint. Notably, Plk1 overexpression increased survival and mutagenesis after Cr(VI) exposure in wild-type Saccharomyces cerevisiae. Taken together, our data indicate that Plk1 activation and nuclear localization are necessary for PTP-regulated mitotic progression after DNA damage. Our studies highlight a role for Plk1 in the loss of checkpoint control, increased survival and mutagenesis after genotoxic exposure in normal cells, which in turn may lead to genomic instability and carcinogenesis.
丝氨酸/苏氨酸激酶 1(Plk1)是有丝分裂的关键调节因子。肿瘤中存在异常的 Plk1 活性,但对于其在正常细胞 DNA 损伤反应中的作用及其对致癌作用早期阶段的潜在贡献知之甚少。DNA 损伤后不适当的存活信号可能促进遗传受损细胞的克隆扩增,并且已知蛋白酪氨酸磷酸酶(PTP)抑制剂可激活关键存活途径。在这项研究中,我们使用六价铬[Cr(VI)],一种有充分文献记载的遗传毒性物质,来研究存活途径激活如何通过涉及 Plk1 的机制导致检查点控制丧失的机制。我们最近报道,PTP 抑制通过覆盖 Cr 诱导的生长停滞,增强 Cr(VI)暴露后的克隆存活和诱变。在这里,我们报告说,PTP 抑制通过 Plk1 活性和核定位的增加来促进检查点绕过,从而促进细胞存活。Plk1 对于正常人类成纤维细胞在 PTP 抑制和 Cr(VI)暴露后的存活增加是必需的,这是通过增强有丝分裂进展来实现的。此外,Plk1 的药理学抑制消除了 PTP 抑制剂诱导的 G(2)/M 检查点绕过。值得注意的是,Plk1 的过表达增加了野生型酿酒酵母在 Cr(VI)暴露后的存活和诱变。总之,我们的数据表明,Plk1 的激活和核定位是 PTP 调节的有丝分裂进展后 DNA 损伤所必需的。我们的研究强调了 Plk1 在正常细胞中遗传毒性暴露后检查点控制丧失、存活增加和诱变中的作用,这反过来可能导致基因组不稳定和致癌作用。