肿瘤特异性survivin启动子在前列腺癌细胞中的活性鉴定

Detection of Transcriptional Activities of Tumor-specific Survivin Promoter in Human Prostatic Carcinoma

目的:克隆survivin启动子片段,并检测其在人前列腺癌LNCaP细胞和人正常张氏肝细胞中的转录活性,为该启动子在前列腺癌的靶向性基因治疗中奠定基础。方法:用PCR扩增survivin基因启动子S1pro和S2pro克隆入pPRIME载体,构建重组质粒pPRIME-S1pro和pPRIME-S2pro,脂质体转染LNCaP细胞和张氏肝细胞,检测survivin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具有较强活性,其中S2pro启动活性明显高于S1pro,达到CMV启动子活性的39%。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 clone DNA sequence of the survivin promoter and study its transcriptional activities in human prostate cancer cells and normal Chang liver cells. Methods： The fragment of the survivin promoter was acquired by PCR amplification and inserted into pPRIME vectors to reconstruct a recombinant plasmid named pPRIME-S1pro and pPRIME-S2pro.Then the reconstructed plasmid was transiently transfected into human prostate cancer cell lines LNCaP and normal Chang liver cells. The transcriptional activities of the survivin promoter in various cells was determined by measuring the expression of green fluorescent protein （GFP）. Results： The survivin promoter had transcriptional activities in LNCaP cells and the transcriptional activity of the S2pro was much higher than that of the S1pro reaching a level of 39% of the transcriptional activity of the CMV promoter. Conclusion ： The survivin promoter cloned in this study is more highly activated in prostate cancer cells than in normal cells. This makes it a potential candidate promoter in the gene therapy for prostate cancer.