Implications of apurinic/apyrimidinic endonuclease in reactive oxygen signaling response after cisplatin treatment of dorsal root ganglion neurons.
Implications of apurinic/apyrimidinic endonuclease in reactive oxygen signaling response after cisplatin treatment of dorsal root ganglion neurons.
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DOI：
10.1158/0008-5472.can-08-1173
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发表时间：
2008-08-01
期刊：
影响因子：
11.2
通讯作者：
Kelley, Mark R.
中科院分区：
文献类型：
--
作者：
Jiang, Yanlin;Guo, Chunlu;Vasko, Michael R.;Kelley, Mark R.
周围神经病变是抗癌药物顺铂的主要副作用之一。尽管先前的研究表明这种神经病变与感觉神经元中DNA加合物的形成有关，但越来越多的证据表明，顺铂也会增加活性氧（ROS）的产生，这可能导致DNA损伤。Ape1/Ref-1是一种多功能蛋白，参与DNA碱基切除修复（BER）和许多转录因子的氧化还原调控。因此，我们询问改变Ape1功能是否会影响顺铂诱导的神经毒性。将培养的感觉神经元暴露于顺铂24小时，并测量几个毒性终点，包括ROS的产生、细胞死亡、凋亡和免疫反应性降钙素基因相关肽（iCGRP）的释放。利用siRNA减少Ape1在神经元培养中的表达，可增强顺铂诱导的细胞杀伤、凋亡、ROS生成以及顺铂诱导的iCGRP释放减少。过表达野生型(WT)-Ape1可减弱顺铂在含有正常内源性Ape1水平的细胞和在ap1sirna处理后Ape1水平降低的细胞中的所有毒性作用。过表达氧化还原缺陷/修复能力的C65-Ape1提供部分拯救，而修复缺陷的Ape1 （N226A+R177A）不能保护神经元免受顺铂毒性。我们还观察到在感觉神经元培养中Ape1水平降低后p53磷酸化增加。这些结果有力地支持了Ape1是一个潜在的翻译靶点的观点，因此保护Ape1水平，特别是其DNA修复功能，可以减少接受顺铂治疗的患者的周围神经病变。
Peripheral neuropathy is one of the major side-effects of the anticancer drug, cisplatin. Although previous work suggests that this neuropathy correlates with formation of DNA adducts in sensory neurons, growing evidence suggests that cisplatin also increases the generation of reactive oxygen species (ROS), which could cause DNA damage. Apurinic/apyrimidinic endonuclease/redox factor-1 (Ape1/Ref-1) is a multifunctional protein involved in DNA base excision repair (BER) of oxidative DNA damage and in redox regulation of a number of transcription factors. Therefore, we asked whether altering Ape1 functions would influence cisplatin induced neurotoxicity. Sensory neurons in culture were exposed to cisplatin for 24 hrs and several endpoints of toxicity were measured including production of ROS, cell death, apoptosis, and release of the immunoreactive calcitonin gene-related peptide (iCGRP). Reducing expression of Ape1 in neuronal cultures using siRNA enhances cisplatin-induced cell killing, apoptosis, ROS generation and the cisplatin-induced reduction in iCGRP release. Overexpressing wild-type (WT)-Ape1 attenuates all the toxic effects of cisplatin in cells containing normal endogenous levels of Ape1 and in cells with reduced Ape1 levels following Ape1siRNA treatment. Overexpressing the redox deficient/repair competent C65-Ape1 provides partial rescue, while the repair deficient Ape1 (N226A+R177A) does not protect neurons from cisplatin toxicity. We also observe an increase in phosphorylation of p53 following a decrease in Ape1 levels in sensory neuronal cultures. These results strongly support the notion that Ape1 is a potential translational target such that protecting Ape1 levels and particularly its DNA repair function could reduce peripheral neuropathy in patients undergoing cisplatin treatment.