Evaluating the role of RAD52 and its interactors as novel potential molecular targets for hepatocellular carcinoma
Evaluating the role of RAD52 and its interactors as novel potential molecular targets for hepatocellular carcinoma
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评估 RAD52 及其相互作用因子作为肝细胞癌新的潜在分子靶标的作用
DOI：
10.1186/s12935-019-0996-6
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发表时间：
2019-11-06
影响因子：
5.8
通讯作者：
Yang, Xiaoli
中科院分区：
文献类型：
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作者：
Li, Ping;Xu, YanZhen;Yang, Xiaoli
背景辐射敏感52（Radiationsensitive 52，RAD 52）是一种重要的肿瘤DNA修复蛋白。然而，关于RAD 52对肝细胞癌（HCC）的影响知之甚少。我们研究了RAD 52在HCC中的表达及其意义。方法应用美国癌症基因组图谱（TCGA）数据库对肝癌组织中的RAD 52 mRNA水平进行分析。采用实时荧光定量PCR（qRT-PCR）、免疫印迹和免疫组化方法检测70例肝癌组织及癌旁组织中RAD 52的表达。研究了在Huh 7 HCC细胞中过表达的RAD 52的作用。然后使用String数据库对RAD 52及其相互作用组进行富集和功能分析。Cytoscape软件用于创建蛋白质-蛋白质相互作用网络。使用Hex8.0.0中的分子对接工具进行与RAD 52及其相互作用组的分子相互作用研究。最后，这些与RAD 52相互作用的DNA修复蛋白也使用TCGA数据集进行了分析，并通过qRT-PCR进行了检测。以TCGA数据库为基础，采用结合RAD 52和RAD 52交互因子的ROC的算法，通过二元logistic regression.ResultsIn TCGA，在HCC中获得与性别相关的RAD 52上调。RAD 52的受试者工作特征曲线（AUC）下的面积为0.704。总生存期（OS）和无复发生存期（RFS）结果表明，RAD 52基因高表达和低表达患者的预后无差异。我们验证了中国人肝癌组织中RAD 52的表达在mRNA和蛋白水平上均高于癌旁组织。较高的RAD 52与年龄相关，与其他临床病理因素无关。在体外，过表达的RAD 52能显著促进Huh 7细胞的增殖和迁移。此外，RAD 52相互作用物（辐射敏感51，RAD 51; X射线修复交叉互补6，XRCC 6; Cofilin，CFL 1）也在HCC中增加，并与RAD 52一起参与一些生物学过程。蛋白质结构分析表明，在RAD 52-RAD 51、RAD 52-CFL 1和RAD 52-XRCC 6复合物中，RAD 52-RAD 51具有最牢固的结合结构，E-总能量最低（− 1120.5 kcal/mol）。RAD 52和其相互作用组之间的ROC相结合的算法表明了更高的特异性和灵敏度HCC screening.ConclusionsOverall，我们的研究表明，RAD 52在HCC发病机制中起着至关重要的作用，并作为一个潜在的分子靶点HCC的诊断和治疗。本研究的结果对于肝癌的多基因预测和诊断具有重要的意义。
BackgroundRadiation sensitive 52 (RAD52) is an important protein that mediates DNA repair in tumors. However, little is known about the impact of RAD52 on hepatocellular carcinoma (HCC). We investigated the expression of RAD52 and its values in HCC. Some proteins that might be coordinated with RAD52 in HCC were also analyzed.MethodsGlobal RAD52 mRNA levels in HCC were assessed using The Cancer Genome Atlas (TCGA) database. RAD52 expression was analyzed in 70 HCC tissues and adjacent tissues by quantitative real-time PCR (qRT-PCR), Western blotting and immunohistochemistry. The effect of over-expressed RAD52 in Huh7 HCC cells was investigated. The String database was then used to perform enrichment and functional analysis of RAD52 and its interactome. Cytoscape software was used to create a protein–protein interaction network. Molecular interaction studies with RAD52 and its interactome were performed using the molecular docking tools in Hex8.0.0. Finally, these DNA repair proteins, which interact with RAD52, were also analyzed using the TCGA dataset and were detected by qRT-PCR. Based on the TCGA database, algorithms combining ROC between RAD52 and RAD52 interactors were used to diagnose HCC by binary logistic regression.ResultsIn TCGA, upregulated RAD52 related to gender was obtained in HCC. The area under the receiver operating characteristic curve (AUC) of RAD52 was 0.704. The results of overall survival (OS) and recurrence-free survival (RFS) indicated no difference in the prognosis between patients with high and low RAD52 gene expression. We validated that RAD52 expression was increased at the mRNA and protein levels in Chinese HCC tissues compared with adjacent tissues. Higher RAD52 was associated with older age, without correlation with other clinicopathological factors. In vitro, over-expressed RAD52 significantly promoted the proliferation and migration of Huh7 cells. Furthermore, RAD52 interactors (radiation sensitive 51, RAD51; X-ray repair cross complementing 6, XRCC6; Cofilin, CFL1) were also increased in HCC and participated in some biological processes with RAD52. Protein structure analysis showed that RAD52–RAD51 had the firmest binding structure with the lowest E-total energy (− 1120.5 kcal/mol) among the RAD52–RAD51, RAD52–CFL1, and RAD52–XRCC6 complexes. An algorithm combining ROC between RAD52 and its interactome indicated a greater specificity and sensitivity for HCC screening.ConclusionsOverall, our study suggested that RAD52 plays a vital role in HCC pathogenesis and serves as a potential molecular target for HCC diagnosis and treatment. This study’s findings regarding the multigene prediction and diagnosis of HCC are valuable.