靶向沉默人HER2/neu基因的RNA干扰  

RNA Interference to Silence Human HER2/neu Gene

作　　者：雷莉妍[1] 孙晓莉[2] 朱利民 修志明[1] 王丽萍[1,2] 

机构地区：[1]吉林大学生命科学学院,长春130012 [2]吉林大学中日联谊医院急救医学科,长春130033 [3]博迅生物技术有限责任公司,长春130012

出　　处：《吉林大学学报（理学版）》2013年第6期1177-1181,共5页Journal of Jilin University:Science Edition

基　　金：吉林省自然科学基金(批准号:201015171);吉林省科技发展计划项目(批准号:201205017);吉林省医药产业发展专项基金(批准号:YYZX201150-2);长春市社会发展科技支撑计划项目(批准号:12SF23)

摘　　要：针对人HER2/neu基因的mRNA序列,设计并构建shRNA干扰载体,经聚合酶链式反应(PCR)及测序鉴定后转染SK-BR-3细胞,并检测HER2/neu mRNA和蛋白抑制情况及SK-BR-3细胞迁移能力的变化.实验结果表明:构建了3个HER2/neu-shRNA真核表达质粒;3个重组质粒均可不同程度下调HER2/neu mRNA和蛋白表达水平,并抑制SK-BR-3细胞的迁移;其中1号重组质粒效应最强,对mRNA和蛋白的抑制率分别为84.65%和74.98%.To construct the short hairpin RNA （shRNA） interference expression plasmid vectors of human HER2/neu gene,the recombinant plasmid was identified by PCR coupled with sequencing.The vector was transfected into SK-BR-3 cells,HER2/neu mRNA,protein,and the cell migration were assayed.The result indicated that the construction of recombinant plasmid was successful.Transfection of shRNA plasmid significantly down-regulated HER2/neu mRNA and protein; at the same time,the migration ability of SK-BR-3 cell was obviously inhibited.Recombinant plasmid 1 had the strongest effect,with the inhibition ratios of 84.65％ at the mRNA level and 74.98％ at the protein level.

关 键 词：HER2 NEU 乳腺癌 shRNA干扰载体 

分 类 号：Q789[生物学—分子生物学]