PSMA增强子/启动子驱动shRNA阻断前列腺癌抗原-1表达治疗前列腺癌的实验研究

项目名称： PSMA增强子/启动子驱动shRNA阻断前列腺癌抗原-1表达治疗前列腺癌的实验研究

项目编号： No.30801151

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： 刘秉乾

作者单位： 郑州大学

项目金额： 22万元

中文摘要： 目前基因治疗前列腺癌在临床I/II期实验效果欠佳，缺乏能用于体内的高效、安全、靶向的基因载体及特异性的靶基因是重要原因。针对以上原因本项目进行了以下研究（1）应用前列腺特异性膜抗原启动子/增强子（PSMAe/p）驱动目的基因表达，成功构建了仅在前列腺癌LNCaP细胞表达的重组质粒；（2）以新近发现的仅在前列腺癌特异表达的PCA-1为靶点，成功构建靶向沉默该基因的重组质粒PSMAe/p-PCA-1-pA-Sliencer，将该质粒转染细胞后可见LNCaP细胞迁移、增殖和浸润生长能力明显降低，其内PCA-1表达明显下降，而PC-3细胞则受影响极小，实现了靶向抑制PCA-1表达；（3）将LNCaP细胞种植到裸鼠皮下以及人的骨片周围成功制备前列腺癌的裸鼠移植瘤和骨转移模型，将转铁蛋白修饰的纳米颗粒载体与重组质粒混合后经静脉注射可见肿瘤生长速度减慢，动物存活时间延长，且肿瘤组织内PCA-1表达下降，对其它主要器官影响较小。我们在国内外首次发现通过靶向PCA-1基因、LNCaP细胞靶向表达和前列腺癌靶向转染的"三重靶向"可以达到抑制前列腺癌发展的目的。

中文关键词： 前列腺癌；前列腺癌抗原-1；基因沉默；前列腺特异性膜抗原

英文摘要： Result of gene therapy is poor for prostate cancer in clinical phase I / II research at present. the lack of efficient, safe, targeted gene vector and specific target gene is one of the important reasons. For the above reasons, this project was carried out following: (1) For expression of target gene was driven by prostate-specific membrane antigen promoter / enhancer (PSMAe/p). we constructed successfully the recombinant plasmid expressing only in prostate cancer LNCaP cells; (2) Due to the PCA-1 gene expressed only in prostate cancer tissue, we constructed successfully the recombinant plasmid PSMAe/p-PCA-1-pA-Sliencer for the PCA-1 as a target. After cells were transfected the recombinant plasmid, migration, proliferation and invasive growth LNCaP cells were reduced significantly, PCA-1 expression was significantly decreased in LNCaP cells, while change of the PC-3 cells is extremely small. We achieved targeted inhibition of the PCA-1 expression; (3) We implant LNCaP cells into skin of nude mice to prepare xenografts model of prostate cancer, and implant LNCaP cells around human bone chip to prepare bone metastasis model of prostate cancer. We found that the expression of PCA-1 can be inhibited, tumor growth be slowed, tumor volume be smaller, survival time of nude mouse be longer and the impact on other major organs be tiny by injection of recombinant plasmid into the xenografts directly or by intravenous injection of mixture of recombinant plasmid and the transferrin-modified nanoparticles vector. By analyzing prostate cancer tissue specimens and follow-up of patients, we found that prognosis be poor in patients with the position of PCA-1 expression. These indicate that the PCA-1 is closely related to prostate cancer. We found that the development of prostate cancer can be inhibited by the "triple targeting" of targeted the PCA-1 gene, targeted expression and targeted transfection in the cellular level and the animal level.

英文关键词： prostate cancer; prostate cancer antigen-1; gene silencing; prostate-specific membrane antigen