去唾液酸糖蛋白受体1（ASGR1）通过Nemo样激酶（NLK）抑制STAT3磷酸化影响肝癌增殖的作用及机制研究

研究肝癌细胞异常增殖的调控机制对肝癌防治具有重要意义。去唾液酸糖蛋白受体1（ASGR1）在正常肝组织中特异高表达，却在肝癌组织中表达沉默，其表达下调的分子机制及下调对肝癌增殖的作用尚未阐明。申请者前期发现：肝癌中ASGR1基因启动子出现高度甲基化，ASGR1低表达和肝癌患者不良预后密切相关；细胞实验发现，ASGR1过表达明显抑制肝癌细胞增殖，且磷酸化的STAT3水平明显降低，提示其通过JAK/STAT3信号通路抑制肝癌细胞增殖。本项目拟在前期研究基础上，通过亲和纯化-质谱技术、ASGR1稳转过表达/沉默肝癌细胞系、Asgr1基因敲除鼠高压注射诱导肝脏原位癌模型、PDX模型，并结合临床样本，从生化、细胞、小鼠、临床患者等层面，深入研究ASGR1在肝癌组织中表达下调的分子机制、其表达下调对肝癌增殖的调节作用、以及通过激活JAK/STAT3信号通路促进肝癌增殖的分子机制，为肝癌的防治提供新思路。

It is of great significance to study the regulatory mechanism of abnormal proliferation for the prevention and treatment of hepatocellular carcinoma (HCC). Asialoglycoprotein receptor 1 (ASGR1) is highly expressed in liver, but it is silenced in HCC. The molecular mechanism of down-regulation of ASGR1 expression and its effect on proliferation of HCC have not been elucidated. The applicant's previous study found that: The promoter of ASGR1 gene was hypermethylated in HCC, and the low expression of ASGR1 was closely related to the poor prognosis of HCC patients. Cell experiment showed that ASGR1 overexpression significantly inhibited the proliferation of hepatoma cells, and the level of phosphorylated STAT3 was significantly decreased, suggesting that ASGR1 could inhibit the proliferation of hepatoma cells through JAK/STAT3 signaling pathway. On the basis of previous studies, this project intends to use co-inmunoprecipitation mass spectrometry technology, ASGR1 overexpression/silencing HCC cell lines, hepatic carcinoma of Asgr1 knockout mice in situ model induced by hydrodynamic injection, PDX model and clinical samples. From the aspects of biochemistry, cell, mice and clinical patients to further study the molecular mechanism of down-regulation of ASGR1 expression in HCC, the regulatory effect on HCC proliferation, and clarify the molecular mechanism of promoting HCC proliferation by activating JAK / STAT3 signaling pathway. This study is intend to provide new ideas for the prevention and treatment of HCC.