TRF2小干扰RNA载体的构建及表达
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宁寒冰, 李继昌, 刘志国, 樊代明
端粒重复序列结合因子2;胃癌;小干扰RNA,宁寒冰,李继昌,刘志国,樊代明,通讯作者:,电话:,收稿日期:,接受日期:,Constructionandexpr
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宁寒冰, 李继昌, 郑州大学第一附属医院消化内科 河南省郑州市 450052
刘志国, 樊代明, 国家肿瘤生物学重点实验室 第四军医大学西京医院全军消化病研究所 陕西省西安市 710032
宁寒冰, 郑州大学医学院博士生, 主要从事消化道肿瘤的研究.
国家自然科学基金资助项目, No. 30400016
通讯作者: 刘志国, 710032, 陕西省西安市长乐西路15号, 第四军医大学西京医院消化病研究所. liuzhiguo@fmmu.edu.cn
电话: 029-84775229
收稿日期: 2006-01-03 接受日期: 2006-01-25
Construction and expression of telomeric repeat binding factor 2 small interfering RNA eukaryotic vector
Han-Bing Ning, Ji-Chang Li, Zhi-Guo Liu, Dai-Ming Fan
Han-Bing Ning, Ji-Chang Li, Department of Digestive Deseases, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China
Zhi-Guo Liu, Dai-Ming Fan, State Key Laboratory of Cancer Biology, Institute of Digestive Diseases, Xijing Hospital, the Fourth Military Medical University, Xi’an 710032, Shaanxi Province, China
Supported by National Natural Science Foundation of China, No. 30400016
Correspondence to: Zhi-Guo Liu, Institute of digestive disease, Xijing Hospital, Fourth Military Medical University, 15 Changle West Road, Xi’an 710032, Shaanxi Province, China. liuzhiguo@fmmu.edu.cn
Received: 2006-01-03 Accepted: 2006-01-25
Abstract
AIM: To construct telomeric repeat binding factor 2 (TRF2) small interfering RNA (siRNA) eukaryotic expression vector and evaluate its expression in gastric cancer cells.
METHODS: Two pairs of hairpin siRNA template oligonucleotides for TRF2 based on pSilencer3.1-H1 vector were designed using the manufacturer’s web-based tool. These oligonucleotides were annealed and ligated into vector respectively. After confirmation by double enzyme digestion analysis and DNA sequencing, positive recombinant plasmids were transfected into multidrug-resistant gastric cancer cells SGC7901/ADR and SGC7901/VCR respectively. Stable cell lines of G418-resistance were acquired after 2-month selection. The effect of transfection on cell growth was analyzed by MTT assay ......
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