前列腺特异性膜抗原启动子调控的重组质粒的构建及表达

目的 探讨前列腺特异性膜抗原(PSMA)启动子调控目的基因表达的前列腺细胞特异性，为前列腺癌靶向性基因治疗作前期铺垫。方法 采用PCR方法从前列腺癌细胞DNA中扩增PSMA启动子序列，将其克隆到含有报告基因——增强绿色荧光蛋白(EGFP)的质粒载体，脂质体介导基因转染不同癌细胞并观察EGFP在不同细胞的表达情况。结果 成功构建质粒pEGFP-PSMAm，质粒转染结果显示PSMA表达阳性的LNeap细胞中存在GFP的有效表达。结论 PSMA启动子调控EGFP表达的重组质粒的构建证明了PSMA启动子的基因转录启动与细胞PSMA的表达相关，即具有前列腺细胞特异性。

The Construction of Recombinant Plasmid with Prostate-specific Membrane Antigen Promoter Controlling Reporter Gene Expression

Objective To study the specificity of prostate-specific membrane antigen(PSMA) promoter in controlling gene expression. Methods PSMA promoter gene was amplified with PCR, and then the promoter gene was cloned into the vector pEGFP-1 to construct a recombinant plasmid, which was transfected into different cell lines such as LNcap, PC-3, MCF-7, A549. Green fluorescent protein(GFP) expression was observed. Results The recombinant plasmid constructed, and PSMA promoter uniquely showed modulating activity in PSMA positive cell line. Conclusion PSMA promoter possesses PSMA positive cell specificity, as well as prostatic tissue specificity. PSMA promoter may have the potential for use in targeted gene therapy of prostate adenocarcinoma.