| DF3启动子调控的hTERT基因RNAi载体的构建及靶向干扰效果的鉴定 |
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| 引用本文： | 曾永秋,赵矫,刘岚,陈绍坤,余红,税青林.DF3启动子调控的hTERT基因RNAi载体的构建及靶向干扰效果的鉴定[J].泸州医学院学报,2011,34(5):521-525. |
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| 作者姓名： | 曾永秋 赵矫 刘岚 陈绍坤 余红 税青林 |
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| 作者单位： | 泸州医学院医学生物学与遗传学教研室,四川泸州,646000 |
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| 基金项目： | 四川省科技厅资助项目(04JY029-020-1) |
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| 摘 要： | 目的：构建带有肿瘤特异性DF3启动子、针对hTERT基因的RNA干扰表达载体pGenesil-10-miR30-DF3-hTERT,探讨该表达载体的肿瘤靶向性基因抑制作用。方法：分别用针对hTERT基因的RNA干扰寡核苷酸序列及DF3启动子取代质粒pGenesil-10-Micro30中原有的miR30序列及CMV启动子,构建pGenesil-10-miR30-DF3-hTERT载体并进行酶切及测序鉴定。将该载体分别转染人乳腺癌MCF-7细胞、肝癌HepG2细胞及造血干细胞,ELISA法检测hTERT蛋白的表达情况。结果：重组质粒经酶切及测序鉴定证实符合设计要求,构建成功。ELISA法检测结果显示,实验组MCF-7细胞及HepG2细胞的hTERT蛋白下降显著（P〈0.05）,其中以MCF-7细胞下降更为明显;造血干细胞实验组则无明显变化（P〉0.05）。结论：DF3启动子调控的hTERT基因的RNA干扰表达载体能有效抑制DF3阳性肿瘤细胞中hTERT的表达,以乳腺癌细胞效果最为显著,而对端粒酶阳性的正常细胞不产生影响。
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| 关 键 词： | DF3启动子 hTERT基因 RNA干扰 肿瘤特异性 |
CONSTRUCTION OF A HTERT GENE-TARGETED RNAI VECTOR REGULATED BY DF3 PROMOTER AND ASSESSMENT OF ITS TARGETED SILENCING EFFECT |
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| Zeng Yongqiu,et al.CONSTRUCTION OF A HTERT GENE-TARGETED RNAI VECTOR REGULATED BY DF3 PROMOTER AND ASSESSMENT OF ITS TARGETED SILENCING EFFECT[J].Journal of Luzhou Medical College,2011,34(5):521-525. |
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| Authors: | Zeng Yongqiu |
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| Institution: | Zeng Yongqiu,et al Department of Medical Biology and Genetics,Luzhou Medical College |
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| Abstract: | Objective: To construct hTERT gene-targeted RNAi expression vector driven by tumor specific DF3 promoter and investigate its tumor-targeted gene suppression effects.Methods: The miR30 sequence and CMV promoter in the plasmid pGenesil-10-Micro30 were replaced respectively by hTERT gene-targeted RNAi oligonucleotides sequence and DF3 promoter.The new recombinant vector pGenesil-10-miR30-DF3-hTERT was confirmed by enzyme digestion and sequencing,and then transfected into three types of cells: human breast canc... |
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| Keywords: | DF3 promoter hTERT gene RNAi Tumor specific |
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