PDCD5蛋白逆转骨肉瘤阿霉素耐药过程中互作蛋白的筛选和功能鉴定

项目名称： PDCD5蛋白逆转骨肉瘤阿霉素耐药过程中互作蛋白的筛选和功能鉴定

项目编号： No.81202124

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： 赵会

作者单位： 首都医科大学

项目金额： 23万元

中文摘要： 骨肉瘤是儿童和青少年最常见的恶性原发骨肿瘤。新辅助化疗和外科手术虽然将五年生存率由10-20%提高到50-60%，但化疗耐药一直是患者术后复发和死亡的主要原因。针对骨肉瘤化疗耐药，目前尚且没有实质性解决方法。PDCD5是北京大学免疫学实验室发现的一种TF1细胞凋亡相关基因，我们首次报道了该基因在体外有效的抑制阿霉素耐药骨肉瘤细胞的增殖，本课题是延伸研究，拟采用串联亲和纯化的方法检测PDCD5分子在耐药骨肉瘤细胞中的蛋白相互作用，分离和鉴定出与PDCD5分子特异性发生作用的蛋白质大分子，验证并确定PDCD5分子与互作蛋白的相互作用结构域，并利用基因功能集合富集度分析方法从基因组水平分析结合蛋白大分子对PDCD5分子转录激活以及细胞整体生物学行为影响。通过对特异性互作蛋白的筛选和研究，为骨肉瘤的耐药发生的分子病因学的研究，也为建立在骨肉瘤胞内信号通路基础上的靶向治疗提供实验基础和理论依据。

中文关键词： 骨肉瘤；PDCD5；蛋白相互作用；串联亲和纯化；

英文摘要： The osteosarcoma of bone is the most common malignant bone tumor among children and adolescents. The five year survival rates have been significant improved from 10-20% to 50-60% since the combination of the neo-adjuvant chemotherapy and surgery. The adjuvant chemotherapeutic resistance is the main reason to recurrence and death for the patients suffered osteosarcoma. But there is no effective method to solve the chemotherapeutic resistance of osteosarcoma. PDCD5 is a novel apoptosis-related gene cloned as an increased expression gene during the apoptotic process of TF-1 cells induced by cytokine withdrawal by Center for Human Disease Genomics of Peking University. We firstly reported that the Adenovirus-delivered PDCD5 could counteract the Adriamycin resistance of osteosarcoma cells through enhancing apoptosis and inhibiting Pgp. This study is the further investigation of the former work. In this research program, we intend to purify the cooperative interactive protein moleculars for PDCD5 protein by the Tandem Affinity Purification (TAP) system, which is manipulated as: constructing a fusion protein of TAP and PDCD5, tranfecting the plasmid to the Saos-2 cells whose PDCD5 is knocked off, purifying the fusion protein by affinity column. Then, the biological attribute of the purified proteins would be identified

英文关键词： osteosarcoma；PDCD5；interactive protein moleculars；tandem affinity purification；