靶向沉默人HER2/neu基因的RNA干扰

吉林大学学报(理学版)

靶向沉默人HER2/neu基因的RNA干扰

雷莉妍¹, 孙晓莉², 朱利民^3, 修志明¹, 王丽萍^1,2

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

收稿日期:2012-12-03 出版日期:2013-11-26 发布日期:2013-11-21
通讯作者: 王丽萍 E-mail:wanglp@jlu.edu.cn

RNA Interference to Silence Human HER2/neu Gene

LEI Liyan1, SUN Xiaoli2, ZHU Limin3, XIU Zhiming1, WANG Liping1,2

1. College of Life Science, Jilin University, Changchun 130012, China；2. Department of Emergency, ChinaJapan Union Hospital, Jilin University, Changchun 130033, China；3. Bioxun Biotech Company Limited, Changchun 130012, China

Received:2012-12-03 Online:2013-11-26 Published:2013-11-21
Contact: WANG Liping E-mail:wanglp@jlu.edu.cn

摘要/Abstract

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%.

关键词: HER2/neu, 乳腺癌； shRNA干扰载体

Abstract:

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.

Key words: HER2/neu; breast cancer, shRNA interfering vector

中图分类号:

Q789

雷莉妍, 孙晓莉, 朱利民, 修志明, 王丽萍. 靶向沉默人HER2/neu基因的RNA干扰[J]. 吉林大学学报(理学版), 2013, 51(06): 1177-1181.

LEI Liyan, SUN Xiaoli, ZHU Limin, XIU Zhiming, WANG Liping. RNA Interference to Silence Human HER2/neu Gene[J]. Journal of Jilin University Science Edition, 2013, 51(06): 1177-1181.