Asparaginyl endopeptidase improves the resistance of microtubule-targeting drugs in gastric cancer through IQGAP1 modulating the EGFR/JNK/ERK signaling pathway.
Asparaginyl endopeptidase improves the resistance of microtubule-targeting drugs in gastric cancer through IQGAP1 modulating the EGFR/JNK/ERK signaling pathway.
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DOI：
10.2147/ott.s125579
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发表时间：
2017
影响因子：
4
通讯作者：
Liu T
中科院分区：
文献类型：
--
作者：
Cui Y;Li Q;Li H;Wang Y;Wang H;Chen W;Zhang S;Cao J;Liu T
近年来，人们对天冬酰胺酰内肽酶（AEP）在肿瘤发生中的作用的了解不断增加。在本研究中，我们研究了胃癌中AEP表达是否与化疗药物敏感性相关，并探讨了其机制。采用酶联免疫吸附法测定患者外周血血清中AEP的表达量。使用单变量和多变量分析评估患者的生存时间。利用质谱分析和免疫共沉淀分析来发现与 AEP 相互作用的蛋白质。建立了胃癌细胞系，其中 AEP 被过表达或使用慢病毒 CRISPR 敲除。使用 Cell Counting Kit-8 方法评估这些细胞系对化疗药物的增殖能力。通过实时聚合酶链反应和蛋白质印迹评估这些品系中的基因表达变化。 AEP低表达的患者在接受多西紫杉醇/S-1方案治疗后明显更有可能具有良好的预后并经历完全缓解或部分缓解。质谱分析表明粘着斑和丝裂原激活蛋白激酶信号通路中的几种蛋白质与 AEP 相互作用。 IQGAP1被证实是与AEP相互作用的蛋白质之一，当AEP被敲除时，其蛋白质水平增加。 AEP 敲除降低了对微管抑制剂的耐药性，包括紫杉醇、多西紫杉醇和 T-DM1。在AEP敲除的胃癌细胞系中，MDR1、p-EGFR、p-JNK、p-ERK和p-Rac1/cdc42的表达水平降低，JNK和ERK抑制剂均可阻断AEP诱导的MDR1表达。 AEP不仅是一个预后因素，也是一个预测标志物。 AEP敲除可以通过与IQGAP1相互作用抑制EGFR/JNK/ERK信号通路的活性并提高对微管抑制剂的敏感性。
In recent years, understanding of the role of asparaginyl endopeptidase (AEP) in tumorigenesis has steadily increased. In this study, we investigated whether AEP expression correlates with sensitivity to chemotherapeutic drugs in gastric cancer and explored the mechanism. AEP expression in the serum of patients’ peripheral blood was measured by enzyme-linked immunosorbent assay. Patient survival time was evaluated using univariate and multivariate analyses. Mass spectrometry and co-immunoprecipitation assays were utilized to discover proteins that interact with AEP. Gastric cancer cell lines were established, in which AEP was overexpressed or knocked out using lentiviral CRISPR. The proliferative abilities of these cell lines in response to chemotherapy agents were evaluated using the Cell Counting Kit-8 method. Gene expression changes in these lines were assessed by real-time polymerase chain reaction and Western blot. Patients with low expression of AEP were significantly more likely to have a good prognosis and experience complete response or partial response after treatment with docetaxel/S-1 regimen. Mass spectrum analysis showed that several proteins in the focal adhesion and mitogen-activated protein kinase signaling pathways interacted with AEP. IQGAP1 was confirmed to be one of the proteins interacting with AEP, and its protein level increased when AEP was knocked out. AEP knockout decreased resistance to microtubule inhibitors, including paclitaxel, docetaxel, and T-DM1. The expression levels of MDR1, p-EGFR, p-JNK, p-ERK, and p-Rac1/cdc42 were decreased in AEP knockout gastric cancer cell lines, and inhibitors of both JNK and ERK could block AEP-induced expression of MDR1. AEP was not only a prognostic factor but also a predictive marker. AEP knockout could inhibit the activity of the EGFR/JNK/ERK signaling pathway and improve sensitivity to microtubule inhibitors through interacting with IQGAP1.