ZBTB16-MTA3/NuRD复合体抑制肝细胞肝癌侵袭与转移的分子机制研究

锌指蛋白在肝癌发生发展过程中扮演重要角色。前期研究发现，ZBTB16在肝癌中低表达且可抑制肝癌侵袭与转移，但分子机制尚不明确。申请者通过蛋白质组学、染色质免疫共沉淀深度测序等技术发现ZBTB16作为MTA3/NuRD复合体新亚基可通过去组蛋白乙酰化修饰抑制RAC1转录。我们推测这可能是ZBTB16抑制肝癌侵袭与转移的分子机制。本研究将从转录因子协同组蛋白乙酰化修饰的角度，进一步探讨ZBTB16-MTA3/NuRD复合体在肝癌发生发展过程中所发挥的作用:首先在分子水平利用FPLC等实验明确该复合体的相互作用机制；然后在转录水平通过qChIP等实验验证该复合体共调控的靶基因；随后在细胞和动物水平研究该复合体对肝癌侵袭与转移的影响及作用机制；最后在临床水平上利用数据库及肝癌组织标本，分析与患者的临床分期、病理分型及预后的关系。本项目旨在为肝癌的研究提供新的线索，并为肝癌的防治提供潜在作用靶点。

Zinc finger proteins play an important role in the progression of hepatocellular carcinoma. It has been implicated that ZBTB16 could suppress HCC invasion and metastasis and the expression of ZBTB16 was lower in tumor tissues than in adjacent tissues. However, the mechanistic role of ZBTB16 in HCC remains an intriguing topic. In light of our previous results, we found that ZBTB16 as a new subunit of MTA3/NuRD complex, might transcriptionally repress RAC1 by physically interacting with HDAC1 through using such technologies as proteomics, ChIP-seq and so on. We speculate that it might be the molecular mechanism that ZBTB16 suppresses HCC invasion and metastasis. In the present study, we will continue working on this complex from the view of transcription factor cooperating with histone deacetylation: firstly, we will focus at the molecular level to study its interaction mechanism by a quantitative approach based on affinity purification-mass spectrometry combined with fast protein liquid chromatography system. Secondly, qChIP will be used to uncover the molecular mechanisms for the target genes regulated by ZBTB16 and MTA3/NuRD at the transcriptional level. Thirdly, molecular, biochemical, cell biological techniques as well as model animal strategy will be further used to reveal the biological significance for the complex. Finally, we will confirm the clinical significance of ZBTB16-MTA3/NuRD-RAC1 in the diagnosis, staging, and prognosis of patients with HCC at the clinical level by TCGA databases, GEO databases and tissues. Over all, our project will elaborate that ZBTB16-MTA3/NuRD complex involved in invasion and metastasis of HCC, providing a molecular mechanism underlying the role of ZBTB16-MTA3/NuRD-RAC1 in invasion and metastasis of HCC and supporting the pursuit of ZBTB16-MTA3/NuRD as potential targets for pharmacological intervention in patients with HCC.