AFP启动子调控PNP载体的构建及分析

目的分别构建三种启动子调控的大肠杆菌嘌呤核苷磷酸化酶(PNP)基因表达载体,检测、分析三者的表达差异.方法构建三种启动子调控的PNP基因表达载体.将三个PNP基因载体转染不同细胞株,检测PNP基因在各细胞株中表达,分析三者表达的特点.结果AF 0.3和[HRE]AF调控的PNP基因在AFP阳性肝癌细胞中实现了靶向性表达,而且后者在AFP阴性肝癌细胞中也实现了靶向性表达.结论三种PNP基因表达载体的成功构建为利用PNP/MeP-dR系统进行肝癌的靶向性基因治疗打下坚实的基础.

Cloning of three vectors carrying PNP gene under the control of different promoters and the analysis of their different expressions

Objective To construct three expression vectors harboring PNP(purine nucleoside phosphorylase) gene under three different promoters .detect and analyze their expressions in different cell lines. Method Three PNP gene expression vectors driven by three different promoters, pcDNA3. 0/PNP, pAF 0. 3/PNP and p[HRE]AF/PNP, were constructed and transfected into four different cell lines, and the expressions of PNP gene in all cell lines were determined. Results The PNP gene expressions of pcDNA3. 0/PNPcould be detected in every cell lines, and the target PNP gene expression of pAF 0. 3/PNP in AFP positive hepatocellular carcinoma cell line was also detected. As for p[HRE]AF/PNP. The target PNP gene expressions were detected both in AFP positive and in negative hepatocellular carcinoma cell lines. Conclusion The successful constructions of three PNP gene vectors will lay a substantial foundation for the target gene therapy of hepatocellular carcinoma with the PNP/ Mep-dR system.