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[稳定的粘着斑激酶基因敲低细胞系的构建及其特性的初步研究]

[Construction of stable focal adhesion kinase knockdown cell line and preliminary study of its properties].

作者信息

Lan Yan, Hua Zi-Chun

机构信息

State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, China.

出版信息

Yao Xue Xue Bao. 2012 Sep;47(9):1128-33.

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.

摘要

恶性黑色素瘤仍然是对健康的严重威胁。黑色素瘤中粘着斑激酶(FAK)的过表达表明,FAK可能是治疗干预的一个有前景的靶点。为了进一步研究FAK在黑色素瘤中的功能,通过将携带FAK小干扰RNA(siRNA)的质粒稳定转染到黑色素瘤细胞系中,下调了FAK的表达。构建并筛选了两个稳定细胞系,F10-siFAK和F10-对照。与F10-对照相比,FAK的mRNA和蛋白水平均显著降低,F10-siFAK的细胞周期停滞在G1期。此外,在体内肿瘤模型中,F10-siFAK细胞的肿瘤生长速度明显慢于F10-对照。这些结果表明,FAK是黑色素瘤中的一个重要调控基因。稳定的FAK敲低黑色素瘤细胞系是进一步研究FAK在黑色素瘤进展中功能的有用工具,也是抗黑色素瘤治疗药物筛选的有效手段。

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