中国普通外科杂志2013，Vol.22Issue(4)：456-462,7.DOI:10.7659/j.issn.1005-6947.2013.04.015
靶向CXCR1基因的shRNA真核表达质粒的构建及鉴定
Construction and identification of eukaryotic expression plasmid encoding shRNA against CXCR1 gene
摘要
Abstract
Objective: To construct the plasmid expression vector that expresses short hairpin RNA (shRNA) against CXC chemokine receptor 1(CXCRl). Methods: Three shRNA expression vectors targeting CXCR1 gene and one negative control (non-targeting sequence) expression vector were designed and constructed according to the mRNA sequence of CXCR1 gene and RNA interference design guidelines. After identification with restriction enzyme digestion and sequencing analysis, these vectors were transfected into gastric cancer MKN45 cells in vitro, and then the CXCR1 mRNA and protein expressions in the cells were detected by RT-PCR and Western blot analysis. Results: As identified by enzyme digestion and sequencing analysis, all the three shRNA eukaryotic expression plasmid vectors targeting CXCR1 gene were successfully constructed. Compared with untransfected MKN45 cells or MKN45 cells transfected with negative control vector, both mRNA and protein levels of CXCR1 were significantly reduced in MKN45 cells transfected with any of the three shRNA vectors (all P<0.05). Conclusion: The successful construction of plasmid expression vector encoding shRNA against CXCR1 may provide a preliminary step for further investigation of the role of CXCR1 in gastric cancer and experimental targeted therapy.关键词
胃肿瘤/受体,CXCR/RNA,小分子干扰Key words
Stomach Neoplasms/ Receptors, CXCR/ RNA, Small Interfering分类
医药卫生引用本文复制引用
王俊普,胡婉明,王宽松,肖燕,李景和,文继舫..靶向CXCR1基因的shRNA真核表达质粒的构建及鉴定[J].中国普通外科杂志,2013,22(4):456-462,7.基金项目
湖南省研究生科研创新资助项目(CX2011B046) （CX2011B046）
湖南省长沙市科技计划资助项目(K1005005-31 （）
K1106041-31). （）
