Integrative proteomics reveals the role of E3 ubiquitin ligase SYVN1 in hepatocellular carcinoma metastasis.
Integrative proteomics reveals the role of E3 ubiquitin ligase SYVN1 in hepatocellular carcinoma metastasis.
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整合蛋白质组学揭示了E3泛素连接酶Syvn1在肝细胞癌转移中的作用。
DOI：
10.1002/cac2.12192
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发表时间：
2021-10
期刊：
影响因子：
--
通讯作者：
Li L
中科院分区：
文献类型：
--
作者：
Ji F;Zhou M;Sun Z;Jiang Z;Zhu H;Xie Z;Ouyang X;Zhang L;Li L
肿瘤转移是肝细胞癌（HCC）预后不良的一个主要因素，但泛素化与转移之间的关系需要更系统地研究。在本研究中，我们分析了HCC的泛素组，以便更全面地了解人类HCC转移情况。
通过泛素组检测了15例有血管侵犯的HCC标本和15例无血管侵犯的HCC标本中的蛋白质泛素化水平。利用在有血管侵犯和无血管侵犯的HCC之间泛素化水平有显著差异的蛋白质来预测与肿瘤转移相关的E3泛素连接酶。构建了蛋白质底物和相应的E3泛素连接酶的拓扑网络以确定关键的E3泛素连接酶。此外，通过细胞增殖实验、皮下肿瘤实验、脐静脉内皮管形成实验、Transwell迁移和侵袭实验，测量了有和无SYVN1表达干扰的LM3和HUH7肝癌细胞系的生长、迁移和侵袭能力。最后，通过蛋白质相互作用组学、免疫荧光和免疫沉淀筛选并验证了SYVN1的相互作用蛋白。比较了阴性对照、SYVN1敲低和SYVN1过表达组中LM3和HUH7细胞中相关蛋白质底物的泛素水平。
在本研究中，我们的全细胞蛋白质组数据集和泛素组数据集包含约5600种蛋白质和12000个泛素化位点。我们发现，在HCC转移过程中，泛素链较短的泛素化位点增加。此外，蛋白质组学和泛素组学分析表明，E3泛素 - 蛋白质连接酶SYVN1的高表达与肿瘤转移有关。进一步，我们发现SYVN1与热休克蛋白90（HSP90）相互作用，并影响真核延伸因子2激酶（EEF2K）的泛素化。
有血管侵犯和无血管侵犯的HCC的泛素化图谱有显著差异。SYVN1是导致这种现象的最重要的E3泛素 - 蛋白质连接酶，且它与肿瘤转移和生长有关。因此，SYVN1可能是HCC的一个潜在治疗靶点。
我们利用高分辨率质谱和二甘氨酸修饰肽的免疫富集构建了迄今为止从人类癌症组织中获得的最大的泛素组数据集之一。我们鉴定出一种E3连接酶SYVN1，它可能参与了HCC的转移。我们研究了SYVN1的相互作用组学，发现它与HSP90相互作用并影响EEF2K的泛素化。
Tumor metastasis is a major factor for poor prognosis of hepatocellular carcinoma (HCC), but the relationship between ubiquitination and metastasis need to be studied more systematically. We analyzed the ubiquitinome of HCC in this study to have a more comprehensive insight into human HCC metastasis. The protein ubiquitination levels in 15 HCC specimens with vascular invasion and 15 without vascular invasion were detected by ubiquitinome. Proteins with significantly different ubiquitination levels between HCCs with and without vascular invasion were used to predict E3 ubiquitin ligases associated with tumor metastasis. The topological network of protein substrates and corresponding E3 ubiquitin ligases was constructed to identify the key E3 ubiquitin ligase. Besides, the growth, migration and invasion ability of LM3 and HUH7 hepatoma cell lines with and without SYVN1 expression interference were measured by cell proliferation assay, subcutaneous tumor assay, umphal vein endothelium tube formation assay, transwell migration and invasion assays. Finally, the interacting proteins of SYVN1 were screened and verified by protein interaction omics, immunofluorescence, and immunoprecipitation. Ubiquitin levels of related protein substrates in LM3 and HUH7 cells were compared in negative control, SYVN1 knockdown, and SYVN1 overexpression groups. In this study, our whole‐cell proteomic dataset and ubiquitinomic dataset contained approximately 5600 proteins and 12,000 ubiquitinated sites. We discovered increased ubiquitinated sites with shorter ubiquitin chains during the progression of HCC metastasis. In addition, proteomic and ubiquitinomic analyses revealed that high expression of E3 ubiquitin‐protein ligase SYVN1 is related with tumor metastasis. Furthermore, we found that SYVN1 interacted with heat shock protein 90 (HSP90) and impacted the ubiquitination of eukaryotic elongation factor 2 kinase (EEF2K). The ubiquitination profiles of HCC with and without vascular invasion were significantly different. SYVN1 was the most important E3 ubiquitin‐protein ligase responsible for this phenomenon, and it was related with tumor metastasis and growth. Therefore, SYVN1 might be a potential therapeutic target for HCC. We constructed one of the largest ubiquitinome dataset obtained from human cancer tissues to date using the high‐resolution mass spectrometry and immunoenrichment of diGly‐modified peptides. We identified a E3 ligase, SYVN1, which potentially participated in the metastasis of HCC. We studied the interaction omics of SYVN1, and found it interacted with HSP90 and impacted the ubiquitination of EEF2K.
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影响因子：
29.4
作者：
Roayaie S;Blume IN;Thung SN;Guido M;Fiel MI;Hiotis S;Labow DM;Llovet JM;Schwartz ME
通讯作者：
Schwartz ME
DNA力学推断的人工端粒形成三链体的动力学
影响因子：
14.9
作者：
Li，Ning;Wang，Junli;Yu，Zhongbo
通讯作者：
Yu，Zhongbo
影响因子：
--
作者：
Kenney, Justin W.;Moore, Claire E.;Proud, Christopher G.
通讯作者：
Proud, Christopher G.
影响因子：
29.4
作者：
Lampis A;Carotenuto P;Vlachogiannis G;Cascione L;Hedayat S;Burke R;Clarke P;Bosma E;Simbolo M;Scarpa A;Yu S;Cole R;Smyth E;Mateos JF;Begum R;Hezelova B;Eltahir Z;Wotherspoon A;Fotiadis N;Bali MA;Nepal C;Khan K;Stubbs M;Hahne JC;Gasparini P;Guzzardo V;Croce CM;Eccles S;Fassan M;Cunningham D;Andersen JB;Workman P;Valeri N;Braconi C
通讯作者：
Braconi C
影响因子：
5.3
作者：
Liu, Liu;Yu, Le;Lin, Zhenghong
通讯作者：
Lin, Zhenghong