| 向 磊,李占魁,邓科委.CMVE-PEG3启动子在前列腺癌DU145细胞中的活性鉴定[J].肿瘤学杂志,2013,19(7):548-552. |
| CMVE-PEG3启动子在前列腺癌DU145细胞中的活性鉴定 |
| Detection of Transcriptional Activities of PEG-3 Promoter Modified by CMV Enhancer in Human Prostate Cancer DU145 Cells |
| 投稿时间：2012-11-30 |
| DOI：10.11735/j.issn.1671-170X.2013.07.B010 |
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| 中文关键词: CMV增强子 PEG-3基因 启动子 DU145细胞 基因治疗 |
| 英文关键词:CMV enhancer PEG-3 promoter DU145 cells gene therapy |
| 基金项目:国家自然科学基金资助项目（30800275） |
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| 中文摘要: |
| 摘 要：［目的］ 比较CMVE-PEG3p、PEG-3启动子在DU145细胞中的启动活性，为前列腺癌靶向性基因治疗提供依据。［方法］ 用PCR法扩增CMV增强子、PEG-3启动子；在真核表达质粒pShuttle-EGFP基础上分别构建2种启动子调控的、以绿色荧光蛋白为目的基因的真核表达质粒pShuttle-CMVE-PEG3p-EGFP、pShuttle-PEG3p-EGFP。将重组质粒用脂质体分别转染前列腺癌DU145细胞和人正常前列腺上皮细胞RWPE-1，72h后用Imagepro-Plus6.0分析2种启动子在相同时间内启动绿色荧光蛋白的表达水平。［结果］ 重组质粒在DU145细胞中观察到了绿色荧光，在RWPE-1细胞中无绿色荧光。pShuttle-CMVE-PEG3p-EGFP质粒在DU145细胞中表达强于pShuttle-PEG3p-EGFP，2种质粒在DU145细胞中的荧光强度(IOD)分别为246.22、130.93。［结论］ 所克隆的CMVE-PEG3p启动子和PEG-3启动子在前列腺癌DU145细胞中均表现出肿瘤特异性，其中CMVE-PEG3p启动子具有更强的启动效应，有望开发成为前列腺癌靶向性基因治疗的工具。 |
| 英文摘要: |
| Abstract：［Purpose］ To compare the transcriptional activities of tumor-specific CMVE-PEG3 promoter and PEG-3 promoter in human prostate cancer DU145 cells，to provide base for targeting gene therapy in human prostate cancer．［Methods］ The fragment of CMVE-PEG3 promoter and PEG-3 promoter were amplified by PCR．The eukaryotic expression plasmids (pShuttle-CMVE-PEG3p-EGFP and pShuttle-PEG3p-EGFP)which have enhanced green fluorescence protein reporter gene and modulated by CMVE-PEG3 promoter and PEG-3 promoter respectively were constructed based on the pShuttle-EGFP．These recombinant plasmids were transfected into DU145 and RWPE-1 cells with liposome．After 72 hours，the expression level of green fluorescent target protein modulated by 2 promoters at the same time was analyzed by Imagepro-Plus6.0 software．［Results］ The green fluorescent target protein was observed in transfected DU145 cells，but there was no green fluorescence in transfected RWPE-1 control cells．The pShuttle-CMVE-PEG3p-EGFP had higher expression activity than pShuttle-PEG3p-EGFP in DU145 cells and the strength rate(IOD) of green fluorescence of DU145 cells transfected by pShuttle-CMVE-PEG3p-EGFP and pShuttle-PEG3p-EGFP were 246.22 and 130.93 respectively．［Conclusion］ Our data reveals that the cloned CMVE-PEG3 and PEG-3 promoter are tumor-specific，and the CMVE-PEG3 promoter has higher transcriptional activities in prostate cancer DU145 cells，and may serve as a useful tool for transcriptional targeting gene therapy of human prostate cancer． |
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