PEG-3启动子在前列腺癌细胞DU145中的活性鉴定  

Activity Identification of PEG-3 Promoter in Prostate Cancer DU145 Cells

作　　者：向磊 李占魁 邓科委 李国权[2] 

机构地区：[1]陕西省妇幼保健院新生儿科,陕西西安710003 [2]第四军医大学西京医院核医学科,陕西西安710032

出　　处：《肿瘤基础与临床》2013年第3期185-188,共4页journal of basic and clinical oncology

基　　金：国家自然科学基金(编号:No.30800275)

摘　　要：目的探讨PEG-3启动子在前列腺癌细胞DU145中的启动活性。方法用PCR法扩增PEG-3启动子;在真核表达质粒pShuttle-CMVp-EGFP基础上构建PEG-3启动子调控的、转录绿色荧光蛋白基因的真核表达质粒pShuttle-PEG3p-EGFP。将2种质粒(pShuttle-PEG3p-EGFP和pShuttle-CMVp-EGFP)用脂质体分别转染前列腺癌细胞DU145和人正常前列腺上皮细胞RWPE-1,72 h后在荧光显微镜下用Im-agepro-Plus 6.0分析2种启动子启动绿色荧光蛋白基因的转录活性。结果重组质粒在前列腺癌细胞DU145中观察到了绿色荧光,在人正常前列腺上皮细胞RWPE-1细胞中无绿色荧光。pShuttle-PEG3p-EGFP和pShuttle-CMVp-EGFP 2种质粒在前列腺癌细胞DU145中的荧光强度分别为403.50、130.93。结论所克隆的PEG-3启动子表现出较强的肿瘤特异性,在前列腺癌细胞DU145中有一定的启动活性,可望开发成为肿瘤靶向性基因治疗工具。Objective To investigate the activities of PEG-3 promoter in prostate cancer DU145 cells. Methods The fragment of PEG-3 promoter was acquired by PCR amplification. The eukaryotic expression plasmids （ pShuttle-PEG3p-EGFP） which had enhanced green fluorescence protein reporter gene and were modulated by PEG-3 promoter were constructed based on the pShuttle-CMVp-EGFP. The two plasmids were transfeeted into the prostate cancer DU145 cells and human normal prostatic epithelium RWPE-1 cells with liposome. After 72 hours, the transcriptional activity of green fluorescent protein modulated by 2 promoters at the same time was analyzed by Imagepro-Plus 6.0 software under the fluorescence microscope. Results The green fluorescent protein was observed in the transfected prostate cancer DU145 cells, but there was no green fluorescence in the transfected prostatic epithelium RWPE-1 control cells. The fluorescence strength of prostate cancer DU145 cells transfected by pShuttle-CM- Vp-EGFP and pShuttle-PEG3p-EGFP were 403.50 and 130.93, respectively. Conclusion The cloned PEG-3 promoter are tumor-specific, and the PEG3 promoter has the transcriptional activities in prostate cancer DU145 cells, and may serve as a useful tool for transcriptional targeting gene therapy of human prostate cancer.

关 键 词：PEG-3 启动子 前列腺癌细胞DU145 基因治疗 

分 类 号：R737.25[医药卫生—肿瘤]