目的:通过研究并比较前列腺特异性膜抗原(PSMA)基因启动子、增强子和survivin基因启动子在不同前列腺癌细胞系(LNCaP和PC-3细胞)中的转录活性,为前列腺癌的靶向性基因治疗提供依据。方法:采用PCR扩增PSMA基因的启动子、增强子和survivin基因启动子,分别克隆入pGL3-Basic,脂质体转染前列腺癌细胞和张氏肝细胞,检测各启动子在细胞中的转录活性。结果:survivin基因启动子在前列腺癌细胞中均具有较强活性,均明显高于PSMA启动子/增强子,其中S2pro启动活性最强,达到CMV启动子活性的1/3,然而,survivin启动子及PSMA启动子/增强子在肝细胞系中几乎不表达。结论:survivin启动子在前列腺癌细胞中具有较强启动活性,可望成为新的前列腺癌靶向性基因治疗工具。
Objective: To detect and compare the transcriptional activities of prostate-specific membrane antigen(PSMA) promoter and enhancer and survivin promoter in different human prostate cancer cell lines,and to search for some evidence for the targeting gene therapy of human prostate cancer.Methods: The fragments of the PSMA promoter and enhancer and survivin promoter were amplified by PCR and inserted into pGL3-Basic.The recombinant plasmids were transiently transfected into human prostate cancer cell lines and normal Chang liver cells,and their transcriptional activities in various cells were determined by measuring the expression of luciferase.Results: The survivin promoter exhibited a higher transcriptional activity than PSMA promoter and enhancer in tumor cell lines,and the S2pro promoter showed the highest activity,reaching one third of that of the CMV promoter.Conclusion: The survivin promoter is highly activated in prostate cancer cell lines and may serve as a new tool for the transcriptional targeting gene therapy of prostate cancer.