泛素化相关蛋白KLHL9 对喉癌增殖和转移的作用及机制

项目名称： 泛素化相关蛋白KLHL9 对喉癌增殖和转移的作用及机制

项目编号： No.81202140

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 肿瘤学2

项目作者： 熊磊

作者单位： 中国人民解放军第二军医大学

项目金额： 23万元

中文摘要： 中晚期喉癌治疗效果仍不理想，原因是喉癌发生发展的机制仍不完全清楚。染色体9p22区域在头颈癌发生发展中经常缺失，我们对这一区域基因做了注释，发现KLHL9是E3连接酶复合物中底物连接蛋白，具有调控细胞分裂的功能，继而我们检测了这个基因在喉癌细胞系和喉癌组织中的表达，发现它在细胞系及癌组织中表达均明显下降，提示该基因可能是抑癌基因，说明泛素化在喉癌发生和发展过程中具有重要作用，但目前尚未见报道。为此，我们将在细胞水平检测该基因对喉癌增殖和转移的影响，并在动物模型中加以验证，并通过临床样本检测，分析KLHL9表达量与喉癌分期、复发、转移和预后的关系；通过酵母双杂交和蛋白质谱技术对KLHL9这个蛋白的底物进行筛选鉴定，利用免疫共沉淀技术验证底物和KLHL9以及泛素的结合，检测底物对喉癌增殖和转移的影响。本研究有望为寻找喉癌新的药物作用靶点和标记物提供理论依据。

中文关键词： KLH9；喉癌；蛋白修饰；抑癌基因；泛素化

英文摘要： The prognosis of laryngeal cancer is poor, which is due to the unknown mechanism of generation and progression of laryngeal cancer. Loss of heterozygosity on chromosome 9p22 frequently take place in head and neck cancer, indicating tumor suppressor genes in this region. To find out the tumor suppressor genes, we analyzed the genes information and found KLHL9, adaptor protein of E3 ligase was capable of regulating cell mitosis. After quantify the expression of KLHL9 by real-time PCR, we found it was less expressed in cancer samples and cancer cell lines, suggesting it is a tumor suppressor. To address the mechanism underlying KLHL9 role on the laryngocarcinogenesis, we will investigate the effect of this gene on the proliferation and metastasis of laryngeal cancer cell lines and verify it in mouse xenograft model. In the meantime, we will also quantify the expression level of KLHL9 in laryngeal cancer tissues and find out its relation to prognosis. Moreover, yeast two hybrid and maldi-TOF will be carried out to fish the substrates of KLHL9 and immunoprecipitation will be used to verify the interaction of substrates with KLHL9 and ubiquitin. The functions of substrates will be investigated in cell lines and mouse xenograft model. This study might provide candidate drug targets and prognosis markers for laryngeal c

英文关键词： KLH9；Laryngeal cancer；protein modification；suppressor genes；polyubiquitinated