生物技术通报Issue(7)：128-133,6.
survivin启动子驱动的shRNA在宫颈癌SiHa细胞中的特异性表达
Specific Expression of shRNA Drived by Survivin Gene Promoter in SiHa Cells
摘要
Abstract
It was to investigate the specific expression of shRNA drived by survivin gene promoter in SiHa cells. The survivin gene promoter was amplified by PCR and pEGFP-C1/Surv plasmid was constructed in which the EGFP was drived by survivin gene promoter, and the activity of survivin gene promoter was detected. shRNA eukaryotic expression vector pGenesil-1-EGFP-shRNA/U6 targeting EGFP was constructed,and the RNA interferencing effect of shRNA drived by U6 promoter was detected. shRNA eukaryotic expression vector pGenesil-1-EGFP-shRNA/Surv in which the U6 promoter was replaced by surviving gene promoter. SiHa and HuVEC cells were transfected by pGenesil-1-EGFP-shRNA/Surv plasmid and the expression of EGFP in two cell lines was observed . Results showed that the survivin gene promoter was amplified successfully and its activity was verified. The pGenesil-1-EGFP-shRNA/U6 plasmid was constructed successfully, and the RNA interferencing effect of shRNA drived by U6promoter was verified as good as expected. The pGenesil-1-EGFP-shRNA/Surv vector was constructed successfully and transfected into SiHa and HuVEC cell lines. After 72 h, there was less green fluorescence in SiHa cells, whereas there was more green fluorescence in HuVEC cells. Therefore,it proved that the survivin gene promoter can drive the specific expression of shRNA in SiHa cell line,which can not happened in normal HuVEC cell line.关键词
survivin启动子/基因治疗/RNA干扰/宫颈癌Key words
Survivin gene promoter Gene therapy RNA interference Cervical cancer分类
医药卫生引用本文复制引用
袁成福,黄秀凝,程莉,卜友泉,刘革力,易发平,杨正梅,宋方洲..survivin启动子驱动的shRNA在宫颈癌SiHa细胞中的特异性表达[J].生物技术通报,2009,(7):128-133,6.基金项目
国家自然科学基金资助项目(30800410,30872758) （30800410,30872758）
