Cullin7泛素连接酶通过NF-κB通路调控CCL2的表达在非小细胞肺癌侵袭转移中的作用机制研究

肺癌是我国发病率和死亡率最高的恶性肿瘤之一。近年许多研究表明单核细胞趋化蛋白-1（CCL2）在肿瘤相关巨噬细胞募集和浸润发挥重要作用。我们研究发现Cullin7（Cul7）泛素连接酶在非小细胞肺癌（NSCLC）组织高表达。通过血管生成因子抗体芯片筛选、Real-time PCR和酶联免疫吸附测定（ELISA）等实验发现Cul7调控CCL2的表达和分泌，并且在NSCLC细胞敲低Cul7能显著逆转TNFα诱导的CCL2表达。本课题拟通过免疫共沉淀(Co-IP)，泛素化降解和免疫荧光等实验阐明Cul7通过NF-κB通路调控CCL2表达的分子机制，并通过CCL2基因敲除小鼠体内肿瘤转移模型和临床肺癌病理组织标本揭示Cul7促进肿瘤侵袭转移的作用。研究结果将为靶向肿瘤相关巨噬细胞治疗和肺癌分子靶向治疗等提供研究思路和关键的实验依据。

Lung cancer is the most common cancer and the leading cause of cancer-related death in China.A lot of studies have shown that monocyte chemotactic protein-1 (CCL2) plays an important role in tumor-associated macrophages recruitment and infiltration. Our previous studies have found that Cul7 ubiquitin ligase is highly expressed in non-small cell lung cancer (NSCLC) tissues. By using antibody microarray screening, enzyme-linked immunosorbent assay (ELISA) and Real-time PCR experiments, we found that Cul7 regulates CCL2 expression. Knockdown of Cul7 can significantly reverse TNFα induced CCL2 expression. The aim of this study is to elucidate the molecular mechanism of Cul7 regulating CCL2 expression via NF-κB signaling by using Co-IP, ubiquitin degradation and immunofluorescence and to explore the role of Cul7 in promoting tumor invasion and metastasis in CCL2 knockout mice tumor metastasis model and clinical lung cancer tissue samples. Our results will provide new research ideas and theoretical evidences for the targeted tumor-associated macrophages treatment of lung cancer.