安徽医科大学学报2012，Vol.47Issue(7)：772-776,5.
真核表达质粒PIRESneo3/miR-194的构建及其在乳腺癌细胞MCF-7中的表达
To construct the eukaryotic expression plasmid PIRESneo3/miR-194 and establish a miR-194 highly expressed MCF-7 cell
摘要
Abstract
Objective To construct the eukaryotic expression plasmid containing miR-194 genes and establish breast cancer cell lines( MCF-7 ) with stabilized expression of miR-194. Methods MiR-194 primers were designed and miR-194 gene was amplified from cDNA of 231 breast cancer cells by reverse transcription polymerase chain re-action ( PCR ). The miR-194 gene was cloned into PIRESneo3 vector directionally after enzyme cut thus constructing the eukaryotic expression plasmid of PIRESneo3/miR-194. Then the recombinant PIRESneo3/miR-194 plasmid was transfected into the MCF-7 cell line by Lipofectamine, and miR-194 highly expressed cells was selected by G418 ( 600 μg/ml ). The expression level of miR-194 was detected by real-time quantitative PCR. Results The recombinant PIRESneo3/miR-194 plasmid was constructed, and the MCF-7 cell line with highly miR-194 gene was screened. Conclusion miR-194 highly expressed MCF-7 cell line is established sucessfully, which will be a perfect target of miR-194 gene therapy and for observing its function in breast cancer cells.关键词
miRNA/转染/乳腺癌/质粒Key words
miRNA/ transfection/ breast cancer/ plasmid分类
医药卫生引用本文复制引用
李佳,王本忠..真核表达质粒PIRESneo3/miR-194的构建及其在乳腺癌细胞MCF-7中的表达[J].安徽医科大学学报,2012,47(7):772-776,5.基金项目
安徽省自然科学基金(编号:11040606M180) （编号:11040606M180）
