PEI载干扰质粒双靶向沉默前列腺癌中核干因子的研究

项目名称： PEI载干扰质粒双靶向沉默前列腺癌中核干因子的研究

项目编号： No.30800226

项目类型： 青年科学基金项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 刘冉录

作者单位： 天津医科大学

项目金额： 21万元

中文摘要： RNA干扰技术的出现为前列腺癌的基因治疗提供了一种新的选择。然而，如果将RNA干扰手段应用于前列腺癌的临床治疗，需要解决两个方面的问题：一是选择能对前列腺癌生长产生显著影响的基因作为干扰的"靶基因"；二是要建立一个安全、高效、特异性强的基因治疗输送系统。在本研究中，课题组针对上述问题，首先利用RNA干扰及基因芯片等相关技术研究表明，核干因子（NS）在前列腺癌中存在高水平表达，而且与前列腺癌的发生发生发展密切相关，将NS 选定为基因治疗的"靶基因"；在此基础上，课题组对原短发夹RNA表达质粒进行改建，成功构建了前列腺特异性膜抗原启动子/增强子（PSMAe/p）作为驱动序列的干扰质粒，在表达水平上靶向前列腺癌，实现了细胞特异性表达，并产生了明显的干扰效果。在载体方面，我们合成并应用聚乙二醇（PEG）修饰的聚乙烯亚胺（PEI）携带特异性干扰质粒，并用转铁蛋白加以修饰，使其对前列腺癌具有靶向性，进行体内外实验，建立了一个安全、高效、特异性强的基因输送及表达系统，为以RNA干扰作为手段的前列腺癌基因治疗奠定了基础。

中文关键词： 前列腺癌；RNA 干扰；核干因子；聚乙烯亚胺；特异性启动子

英文摘要： RNA interferenec(RNAi) provides a new choice for the treatment of prostate cancer. However, two problems must be resolved if RNAi could be used for the therapy of prostate cancer clinically: one is that a suitable gene must be chosen as the "target gene", the other is to establish a safe, effective and specific gene transfer system. To resolve the two problems, in this study, we first chose Nucleostemin(NS) as the "target gene" because its high expression level in prostate cancer and its tight correlation with the initiation and development of prostate cancer by RNAi and genechips. Based on these results, we rebuilt the short-hair RNA expressing plasmid and PSMA promoter/enhancer was used as the driving sequence to tatget the prostate cancer at the expression level and accomplished cell-specific expression. About the vector, polyethylene glycol-modified polyethyleneimine was synthesized and used to transfer the plasmid, and transferrin was linked to the vector to target prostate cancer. In the end, a safe, effective and specific gene transfer and expression system was established by ex vivo and in vivo experiment. Our study established a solid basis for the gene therapy of prostate caner by using RNAi.

英文关键词： prostate cancer;RNA interference; Nucleostemin;polyethyleneimine;specific promoter