WD repeat-containing protein 1 maintains β-Catenin activity to promote pancreatic cancer aggressiveness
WD repeat-containing protein 1 maintains β-Catenin activity to promote pancreatic cancer aggressiveness
复制标题
含有 WD 重复序列的蛋白 1 保持 β-连环蛋白活性，促进胰腺癌侵袭性
DOI：
10.1038/s41416-020-0929-0
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发表时间：
2020-06-30
影响因子：
8.8
通讯作者：
Fu, Deliang
中科院分区：
文献类型：
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作者：
Li, Hengchao;Liu, Xiaohui;Fu, Deliang
背景胰腺导管腺癌(PDAC)进展的分子特征可能包括影响恶性表型的关键蛋白。在这里，我们的目标是确定与PDAC相关的不同肿瘤转移(TNM)阶段的蛋白。方法采用8组等压标签结合二维液相色谱-串联质谱仪分析不同TNM分期的PDAC组织的蛋白质组。通过功能丧失研究来评估WD重复蛋白1(WDR1)在PDAC中的致癌作用。采用实时荧光定量聚合酶链式反应、Western blotting、邻近连接实验和免疫共沉淀等方法研究了WDR1促进PDAC进展的分子机制。结果共鉴定出5036个蛋白质组分，4708个蛋白质组分的可信度较高。与正常胰腺组织相比，PDAC各期组织中37种蛋白均发生了显著变化。此外，随着肿瘤TNM分期的增加，蛋白表达上调或下调，有10个蛋白与肿瘤的发生有关。功能不明的WDR1蛋白在PDAC中高表达，预示预后不良。WDR1基因敲除抑制了PDAC肿瘤的生长和体内转移。此外，WDR1基因敲除抑制了Wnt/β-catenin途径的活性；异位表达稳定形式的β-catenin恢复了WDR1基因敲除的抑制作用。在机制上，WDR1与USP7相互作用，以阻止泛素化介导的β-连环蛋白的降解。结论我们的研究发现了几个与PDAC进展有关的未知功能蛋白，并为WDR1在PDAC发生发展中的致癌作用提供了新的见解。
Background The molecular signature underlying pancreatic ductal adenocarcinoma (PDAC) progression may include key proteins affecting the malignant phenotypes. Here, we aimed to identify the proteins implicated in PDAC with different tumour-node-metastasis (TNM) stages. Methods Eight-plex isobaric tags coupled with two-dimensional liquid chromatography-tandem mass spectrometry were used to analyse the proteome of PDAC tissues with different TNM stages. A loss-of-function study was performed to evaluate the oncogenic roles of WD repeat-containing protein 1 (WDR1) in PDAC. The molecular mechanism by which WDR1 promotes PDAC progression was studied by real-time qPCR, Western blotting, proximity ligation assay and co-immunoprecipitation. Results A total of 5036 proteins were identified, and 4708 proteins were quantified with high confidence. Compared with normal pancreatic tissues, 37 proteins were changed significantly in PDAC tissues of different stages. Moreover, 64 proteins were upregulated or downregulated in a stepwise manner as the TNM stages of PDAC increased, and 10 proteins were related to tumorigenesis. The functionally uncharacterised protein, WDR1, was highly expressed in PDAC and predicted a poor prognosis. WDR1 knockdown suppressed PDAC tumour growth and metastasis in vitro and in vivo. Moreover, WDR1 knockdown repressed the activity of the Wnt/beta-Catenin pathway; ectopic expression of a stabilised form of beta-Catenin restored the suppressive effects of WDR1 knockdown. Mechanistically, WDR1 interacted with USP7 to prevent ubiquitination-mediated degradation of beta-Catenin. Conclusion Our study identifies several previous functional unknown proteins implicated in the progression of PDAC, and provides new insight into the oncogenic roles of WDR1 in PDAC development.