Construction of stable focal adhesion kinase knockdown cell line and preliminary study of its properties.
- Author:
Yan LAN
1
;
Zi-Chun HUA
Author Information
1. State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Cell Cycle;
Cell Line, Tumor;
Cell Proliferation;
Down-Regulation;
Focal Adhesion Protein-Tyrosine Kinases;
genetics;
metabolism;
G1 Phase;
Gene Knockdown Techniques;
Melanoma, Experimental;
enzymology;
pathology;
Mice;
Mice, Inbred C57BL;
Mitogen-Activated Protein Kinase 1;
metabolism;
Mitogen-Activated Protein Kinase 3;
metabolism;
Plasmids;
RNA, Messenger;
metabolism;
RNA, Small Interfering;
genetics;
Transfection
- From:
Acta Pharmaceutica Sinica
2012;47(9):1128-1133
- CountryChina
- Language:Chinese
-
Abstract:
Malignant melanoma still remains to be a serious health threat. Overexpression of focal adhesion kinase (FAK) in melanoma has suggested that FAK could be a promising target for therapeutic intervention. To further investigate the function of FAK in melanoma, FAK expression was down-regulated by stable transfection of plasmid harboring FAK small interfering RNA (siRNA) into melanoma cell line. Two stable cell lines, F10-siFAK and F10-control, have been constructed and screened. Compared with the F10-control, both the mRNA and protein levels of FAK decreased significantly, and the cell cycle of F10-siFAK was arrested at G1 phase. Furthermore, the tumor growth rate of F10-siFAK cells was notably slower than that of F10-control in in vivo tumor models. These results show that FAK is an important regulatory gene in melanoma. The stable FAK-knockdown melanoma cell line is an useful tool for further investigation of FAK's function in the progression of melanoma, and also an effective means of drug screening for anti-melanoma therapeutics.
