Proteomic analyses reveal divergent ubiquitylation patterns in hepatocellula carcinoma cell lines with different metastasis potential
Proteomic analyses reveal divergent ubiquitylation patterns in hepatocellula carcinoma cell lines with different metastasis potential
复制标题
蛋白质组学分析揭示具有不同转移潜力的肝细胞癌细胞系中不同的泛素化模式
DOI：
10.1016/j.jprot.2020.103834
复制
发表时间：
2020-04-15
影响因子：
3.3
通讯作者：
Zhao，Bixing
中科院分区：
文献类型：
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作者：
Sun，Ying;Zheng，Xiaoyuan;Zhao，Bixing
肝细胞癌（Hepatocellular carcinoma，HCC）是最常见的恶性肿瘤之一，转移和复发是影响预后的主要原因。泛素化作为蛋白质的降解机制，但它涉及其他细胞过程，包括转移。在这里，我们通过使用无标记定量，双甘氨酸（di-Gly）抗体亲和纯化和高分辨率液相色谱串联质谱（LC-MS/MS），研究定量蛋白质组，泛素组，以及两个数据集之间的串扰在不同转移潜力的肝癌细胞系，以确定与肝癌转移相关的生物标志物。总共有83个泛素化蛋白质根据其转移潜力显著而稳定地改变其丰度，并且这些泛素化蛋白质参与的生物学过程与肿瘤转移密切相关。进一步的信号通路分析显示，在高转移细胞中，核糖体和蛋白酶体被显著过度激活。此外，我们还分析了整个蛋白质组与泛素化组之间的相互作用，并进一步探讨了泛素化事件影响肝癌转移的机制。最终，Ku 80的泛素化被证实在高转移细胞中与低转移细胞相比显著下调。我们相信这些发现将有助于我们更好地了解HCC转移的潜在分子机制。Significance在这篇文章中，我们使用基于无标记的蛋白质组学结合双甘氨酸抗体（di-Gly）亲和纯化方法，在一系列具有增加侵袭和转移潜力的HCC细胞系中鉴定与HCC复发/转移相关的生物标志物。然后，我们分析了整个蛋白质组和泛素组之间的串扰。最终证实Ku 80的泛素化与肝癌细胞的侵袭和迁移密切相关。据我们所知，这是第一次利用定量蛋白质组学的方法来研究具有转移能力的肝癌细胞系中的泛素组学。
Hepatocellular carcinoma (HCC) is one of the most common malignant tumours, metastasis and recurrence remain the primary reasons for poor prognosis. Ubiquitination serves as a degradation mechanism of proteins, but it is involved in additional cellular processes including metastasis. Here, by using label-free quantification, double-glycine (di-Gly) antibody affinity purification and high-resolution liquid chromatography tandem mass spectrometry (LC-MS/MS), we investigated quantitative proteome, ubiquitylome, and the crosstalk between the two datasets in HCC cell lines with different metastasis potential to identify biomarkers associated with HCC metastasis. In total, 83 ubiquitinated proteins significantly and steadily changed their abundance according to their metastatic potential, and the participated biological processes of these ubiquitinated proteins were tightly associated with tumour metastasis. Further signaling pathway analysis revealed that the ribosome and proteasome were significantly over-activated in the highly metastatic cells. Furthermore, we analyzed the crosstalk between the whole proteome and the ubiquitylome, and further discussed the mechanism that how ubiquitination events affect HCC metastasis. Eventually, the ubiquitination of Ku80 was validated to be significantly down-regulated in the high-metastatic cells comparing with the low-metastatic cells. We believe that these findings will help us better understand the underlying molecular mechanisms of the metastasis of HCC.SignificanceIn this manuscript, we used label free based proteomics combined with diglycine antibody (di-Gly) affinity purification approach to identify biomarkers associated with HCC recurrence/metastasis in in a serial HCC cell lines with increasing invasion and metastasis potential. And then, we analyzed the crosstalk between the whole proteome and the ubiquitylome. Eventually, the ubiquitination of Ku80 was confirm to be closely associated with invasion and migration of HCC cells. As far as we know, this is the first time to use quantitative proteomic approach to study the ubiquitylomics in HCC cell lines with increasing metastasis ability.