一种新型诱导细胞凋亡重组蛋白的研制以及与抗新生血管生成协同治疗肝癌的研究

项目名称： 一种新型诱导细胞凋亡重组蛋白的研制以及与抗新生血管生成协同治疗肝癌的研究

项目编号： No.30872987

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 金属学与金属工艺

项目作者： 孙学英

作者单位： 哈尔滨医科大学

项目金额： 34万元

中文摘要： 肝细胞肝癌是我国高发疾病，Survivin是肝癌细胞生存所必须的因子，本研究设计合成了一种新型的Survivin显性失活突变体重组蛋白命名为R9DnSur。体外实验发现R9DnSur可以快速进入肝癌细胞诱导细胞凋亡和抑制细胞增殖。但是这种重组蛋白对于非Survivin依赖性的其他肿瘤细胞和血管内皮细胞没有影响，说明它对肿瘤新生血管没有作用。本课题联合应用R9DnSur和抗新生血管生成剂，索拉菲尼，进行肝癌治疗的新探讨。 联合应用的理论基础是两种治疗方法同时针对肝癌的不同细胞群，即肝癌细胞和血管内皮细胞。 项目还对基因表达水平、肿瘤细胞凋亡通路、肿瘤血管化的程度等进行检测，从分子水平探讨这种治疗方法的机理。项目对如下关键性问题进行探讨：1) R9DnSur重组蛋白合成和纯化以及大量制备；2)R9DnSur重组蛋白体外功能的检测和分子通路; 3)裸鼠人肝癌模型的建立（包括局部结节性肝癌和弥漫性肝癌）4）R9DnSur重组蛋白的剂量和时间的选择；5)R9DnSur与索拉菲尼联合作用的机理和对肝癌细胞分子信号通路的影响。本研究提示R9DnSur重组蛋白可有效提高索拉菲尼治疗肝癌的效果。

中文关键词： 生存素；凋亡;重组蛋白；肝癌；新生血管生成

英文摘要： Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide, particularly in china. It is also the second leading cause of cancer-related deaths among men worldwide. Survivin is an important survival factor for HCC. Several approaches have been launched by targeting surviving to combat cancer. In this project, we have designed a novel recombinant dominant negative protein targeting survivin, defined as R9DnSur, which could quickly penetrate the cells, and inhibit the proliferation and induce apoptosis of HCC cells, but has no effects on surviving-independent cells, such as vascular endothelial cells, indicating it has no effect on tumor angiogenesis. Here, we have combined R9DnSur protein therapy with anti-angiogenic agent, and test their synergistic effect to treat HCC in mice. R9DnSur and an anti-angiogenic agent, sorafenib, inhibited tumor growth by themselves and synergized to eradicate established xenografts of HepG2 tumors in mice. We have also investigated the anti-tumor mechanisms at molecular levels including gene expression, cell death signaling pathways and vascularization degree. The present study focuses on the following key points: 1) synthesis, purification and preparation of recombinant protein R9DnSur; 2) extrinsic functional monitoring and molecular pathways of recombinant protein R9DnSur; 3) subcutaneous establishment of HepG2 xenografts in the mice; 4) dosage and administration routs of R9DnSur; 5) synergy of R9DnSur and sorafenib. A conclusion from this study has been drawn that recombinant protein R9DnSur warrants investigation as a potential strategy to enhance the efficacy of sorafenib for treating HCC. Under the financial support of NNSF (30872987), the research group has collaborated with other oversea researchers and utilized financial supports from other funding bodies, in investigating cell apoptosis, hypoxia microenvironments, angiogenesis, and cell signaling pathways of HCC. Up to December, 2011, 21 research articles have been published in SCI cited peer-viewed journals, and 5 postgraduate students trained for their Doctor's Degrees.

英文关键词： survivin; apoptosis; recombinant protein;hepatocellular carcinoma;angiogenesis