RNA干扰体外抑制人食管鳞癌细胞Eca-109缺氧诱导因子-1α的表达
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肖 斌, 施瑞华, 杜琰萍, 朱 宏, 凌亭生,
食管鳞癌;缺氧诱导因子-1α;RNA干扰;短发夹状RNA,肖斌,施瑞华,杜琰萍,朱宏,凌亭生,张国新,林艳,郝波,肖斌,通讯作者
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肖斌, 施瑞华, 杜琰萍, 朱宏, 凌亭生, 张国新, 林艳, 郝波, 南京医科大学第一附属医院消化科 江苏省南京市 210029
肖斌, 1997年南京医科大学本科毕业, 主治医师, 硕士在读, 主要从事消化系统疾病的临床与基础研究.
江苏省自然科学基金资助项目, No. BK2005153
通讯作者: 施瑞华, 210029, 江苏省南京市广州路300号, 南京医科大学第一附属医院消化科. ruihuashi@126.com
电话: 025-83718836-6973
收稿日期: 2006-03-24 接受日期: 2006-05-09
RNA interference inhibits expression of hypoxia inducible factor-1α in human esophageal squamous cancer cell line Eca-109 in vitro
Bin Xiao, Rui-Hua Shi, Yan-Ping Du, Hong Zhu, Ting-Sheng Ling, Guo-Xin Zhang, Yan Lin, Bo Hao
Bin Xiao, Rui-Hua Shi, Yan-Ping Du, Hong Zhu, Ting-Sheng Ling, Guo-Xin Zhang, Yan Lin, Bo Hao, Department of Gastroenterology, the Frist Affiliated Hospital of Nanjing Medical University, Nanjing 210029, Jiangsu Province, China
Supported by the Natural Science Foundation of Jiangsu Province, No. BK2005153
Correspondence to: Dr. Rui-hua Shi, Department of Gastroenterology, the Frist Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing 210029, Jiangsu Province, China. ruihuashi@126.com
Received: 2006-03-24 Accepted: 2006-05-09
Abstract
AIM: To investigate inhibitory effect of hypoxia inducible factor-1α (HIF-1α) short hairpin RNA (shRNA) on the expression of HIF-1α in human esophageal squamous cancer cell line Eca-109, and to screen suitable interference targets in HIF-1α gene.
METHODS: Three pairs of oligonucleotides targeting HIF-1α were cloned into linearized PGCsi eukaryotic expression vector after annealing. The recombinant plasmids were identified by sequencing. Human esophageal squamous cancer cells Eca-109 and embryonic kidney cells 293T were transfected with the recombinant plasmids, respectively. Fluorescence microscopy was used to observe the effect of RNA interference and evaluate the efficacy of transfection. The transcription of HIF-1α gene and the expression of HIF-1α protein were determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, respectively ......
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