Construction of the siRNA expression vector and the silencing influence on focal adhesion kinase gene of Tca8113 cell line
- VernacularTitle:靶向黏着斑激酶siRNA表达载体构建及转染Tca8113细胞的沉默效应
- Author:
Hualian LIU
;
Hongbing JIANG
;
Shuzhong XING
;
Laikui LIU
;
Zilu WANG
;
Yangyu ZHENG
- Publication Type:Journal Article
- Keywords:
Focal adhesion kinase;
Tongue cancer cell;
RNA interference
- From:
Journal of Practical Stomatology
2001;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct the siRNA expression vector of focal adhesion kinase(FAK) gene and inhibit the expression of FAK gene in tongue cancer cell line Tca8113 by RNA interfering technique. Methods:According to the encoding sequence of FAK mRNA, two pieces of oligonucleotide sequences were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pGCSilencerTM-U6/Neo siRNA expression vector. After being identified by restriction enzyme method, the recombinant pSilencer-FAK plasmids were transfected into Tca8113 cells. The transfected cells were selected by G418 method. Immuocytochemistry and Western blotting were used to evaluate FAK gene silencing efficiency. Results:The oligonucleotide fragments were correctly inserted into pGCSilencerTM-U6/Neo vector. FAK expression of the transfected cells was significantly down-regulated by pSilencer-FAK. Conclusion:The siRNA expression vector of FAK is successfully constructed and FAK expression of Tca8113 cells can be inhibited by RNA interfering technique.
