浙江医学Issue(9)：747-749,761,4.
Livin shRNA 表达载体的构建及在胰腺癌细胞的效应研究
Construction of LivinshRNA vector and its effect on expression of Livin mRNA and protein in pancreatic cancer cells
摘要
Abstract
Objective To construct LivinshRNA eukaryotic expression vectors and to investigate its effect on ex-pression of Livin mRNA and protein in pancreatic cancer cel line panc- 1. Methods Livin gene interference sequence was synthesized and inserted into pGCsilencerTM U6/Neo/GFP/RNAi vector,which was confirmed by sequencing. The recombi-nant RNAi vector was transfected into pancreatic cancer panc- 1cel s by Lipofectamine 2000. The cel s containing stable transformants were selected by G418, and isolated with limited dilution. The mRNA expression of Livin in the selected clones was detected by RT- PCR, the protein expression of Livin was detected by Western blot. Results The successful construction of RNAi eukaryotic expression vectors targeting Livin was confirmed by sequencing, which was expressed in panc- 1 cel s after transfection as showed by green fluorescence under inverted fluorescence microscope. RT- PCR and West-ern blot revealed that the expression of Livin mRNA and protein was down- regulated in panc- 1 cel s. Conclusion Livin shRNA eukaryotie expression vector was successful y constructed and the transfected pancreatic cancer panc- 1 cel s show a down- regulated expression of Livin mRNA and protein.关键词
Livin基因/RNA干扰/转染/panc-1细胞Key words
Livin gene/RNA interference/Transfection/panc- 1 cell引用本文复制引用
张财明,季一鸣,朱敏,吕尚东,王爱东..Livin shRNA 表达载体的构建及在胰腺癌细胞的效应研究[J].浙江医学,2015,(9):747-749,761,4.基金项目
台州市科技局计划项目 （）
