小干扰RNA抑制胃癌细胞株葡萄糖调节蛋白Grp78的表达
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张新晨, 杨维良, 徐华锋, 吴德全, 杨宝峰
胃癌;RNA干扰;葡萄糖调节蛋白;逆转录-聚合酶链反应;免疫荧光技术,张新晨,杨维良,吴德全,徐华锋,杨宝峰,张新晨,通讯作者:,RepressiveeffectsofsmallinterferingRNAonexpressio
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张新晨, 杨维良, 吴德全, 哈尔滨医科大学附属第二医院普通外科 黑龙江省哈尔滨市 150086
徐华锋, 黑龙江大学生命科学院 黑龙江省哈尔滨市 150080
杨宝峰, 哈尔滨医科大学药理学教研室 150080
张新晨, 副主任医师, 医学博士, 博士后, 主要从事普通外科、肿瘤基因治疗方面研究.
黑龙江省自然科学基金资助项目, No. D2004-23
通讯作者: 杨宝峰, 150080, 黑龙江省哈尔滨市, 哈尔滨医科大学药理学教研室. yangbf@ems.hrbmu.edu.cn
电话: 0451-86605356
收稿日期: 2006-10-31 接受日期: 2006-12-14
Repressive effects of small interfering RNA on expression of glucose-regulated protein Grp78 in human gastric cell line
Xin-Chen Zhang, Wei-Liang Yang, Hua-Feng Xu, De-Quan Wu, Bao-Feng Yang
Xin-Chen Zhang, Wei-Liang Yang, De-Quan Wu,Department of General Surgery, the Second Affiliated Hospital of Harbin Medical University, Harbin 150086, Heilongjiang Province, China
Hua-Feng Xu,College of Life Science, Heilongjiang University, Harbin 150080, Heilongjiang Province, China
Bao-Feng Yang, Department of Pharmacology, Harbin Medical University, Harbin 150086, Heilongjiang Province, China
Supported bythe Natural Science Foundation of Heilongjiang Province, No. D2004-23
Correspondence to:Professor Bao-Feng Yang, Department of Pharmacology, Harbin Medical University, Harbin 150086, China. yangbf@ems.hrbmu.edu.cn
Received:2006-10-31 Accepted:2006-12-14
Abstract
AIM: To construct the eukaryotic expression vector of RNA interference specific for glucose-regulated protein Grp78, and to observe silencing effect on Grp78 expression in human gastric cell line 7901.
METHODS: Gene sequences of human Grp78 were obtained from GenBank. A couple of Grp78 gene hairpin oligonucleotide was designed, whose sequence met the need of psiSTRIKETM U6 small interfering RNA (siRNA) expression vector. The designed oligonucleotide was inserted into plasmid psiSTRIKETM, which was an eukaryotic expression vector controlled by the U6 promoter of RNA polymerase Ⅲ, and then human gastric cell line 7901 was transfected with the recombinant plasmid by Lipofectamin2000 ......
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