RNAi technology to block the expression of molecules relevant to metastasis: the cell adhesion molecule CEACAM1 as an instructive example.
Specific gene silencing using small hairpin RNA (shRNA) constructs offer researchers the possibility to study the influence of a single protein in the metastatic process. The role of the cellular adhesion molecule CEACAM1 on tumour formation and metastasis is of some interest. The human melanoma cell line FemX-1 was transfected with an shRNA construct directed against the CEACAM family. Stable clones were obtained and characterized via puromycin selection, single-cell dilution, and subsequent FACS analysis. The cell line showed a knock-down of CEACAM1 of more than 85%. This knock-down remained stable when examined in an SCID mouse xenograft experiment over 40 days.
Centre for Experimental Medicine, Institute for Anatomy and Experimental Morphology, Hamburg, Germany. d.wicklein@uke.uni-hamburg.de
Daniel
D
Wicklein
D015703:Antigens, CD / Q000235:genetics*; D015815:Cell Adhesion Molecules / Q000172:deficiency* / Q000235:genetics*; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D055785:Gene Knockdown Techniques; D005821:Genetic Techniques; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D009362:Neoplasm Metastasis / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection
D015703:Antigens, CD; C069847:CD66 antigens; D015815:Cell Adhesion Molecules; D034741:RNA, Small Interfering