项目名称: HDAC2磷酸化激活自身SUMO E3连接酶活性促进结肠癌细胞生长的机制研究
项目编号: No.81502434
项目类型: 青年科学基金项目
立项/批准年度: 2016
项目学科: 医药、卫生
项目作者: 邢伟
作者单位: 中国人民解放军第三军医大学
项目金额: 16.5万元
中文摘要: HDAC2在肿瘤发生和发展中起重要作用,是有效的抗癌靶点。我们前期研究报道,HDAC2还兼有独立的SUMO E3连接酶功能域,此功能介导调控与细胞增殖和抗凋亡相关基因的蛋白质翻译水平表达,在维持结肠癌细胞快速生长中发挥重要作用。另发现HDAC2 T480潜在的磷酸化修饰是激活自身SUMO E3连接酶活性的关键,而其中的机制仍不清楚。为此本研究将利用质谱、磷酸化抗体技术进一步鉴定T480的磷酸化及其蛋白激酶;利用SUMO化、Pull-down等实验方法,研究HDAC2T480A突变体对自身及其靶蛋白eIF4E的SUMO化和对翻译起始复合物eIF4F的影响及调控机制;检测c-myc、bcl2、survivin等靶基因的蛋白质新生速率;突变体抑制裸鼠成瘤的作用。揭示HDAC2 T480磷酸化激活自身SUMO E3连接酶活性促进结肠癌细胞生长的机制,为进一步设计研发靶向此连接酶的抗癌新药奠定基础。
中文关键词: C08_结;直肠肿瘤;组蛋白去乙酰化酶2;磷酸化;小泛素样修饰
英文摘要: HDAC2 has an important role in the tumorigenesis and is an effective anti-tumor target. Our previous reports shown that HDAC2 had an independent SUMO E3 ligase domain, which mediated the translation control of genes associated with proliferation, growth and anti-apoptosis, and played an important part in cancer cell growth. However, in the preliminary experiments, we found that the potential phosphorylation of threonine T480, the 480th amino-acid residue in the C-terminal of HDAC2, was the key event for activating the SUMO-E3 ligase activity of HDAC2. But the involved molecular mechanism remains to be explored. By using LC-MS and antibody for detecting phosphorylated protein, we will further determine the phosphorylation of T480 threonine residue and its protein kinase. By using SUMO-modification and Pull-down technique, we will research the effect of HDAC2T480A mutant on the SUMO-modification of HDAC2 and eIF4E, and the effect on translational initial complex 4F(eIF4F). we will determine the effect of T480A mutant on the protein biosynthesis rate of target genes such as c-myc, bcl2 and survivin. Furthermore, we will research the anti-oncogenic characteristic of T480A mutant in nude mouse. Eventually, we will reveal the mechanism that HDAC2 T480 phosphorylation stimulates itself SUMO E3 ligase activity. The project will facilitate the further design and development of new anticancer drug targeted to the SUMO E3 ligase.
英文关键词: colorectal cancer;HDAC2; phosphorylation;SUMO