C-Flipp43 Induces Activation Of The Nuclear Factor-Kappa B Signaling Pathway In A Dose-Dependent Manner In The A375 Melanoma Cell Line

In order to investigate the role of c-FLIPp43 in the regulation of the nuclear factor (NF)-kappa B signaling pathway in melanoma cell lines, a eukaryotic expression vector for c-FLIPp43 was constructed with the pCMV-Tag2B plasmid. The monoclonal A375 cells with stable expression of c-FLIPp43 were obtained by G418 selection and were identified with western blot analysis. The protein level of NF-kappa B-p65 in the A375 cell line with stable expression of c-FLIPp43 was examined by western blot analysis. The translocation of NF-kappa B-p65 was examined using immunofluorescence. The A375 cell lines were transfected with the pCMV-Tag2B-cFLIP(p43) vector at different doses and the activation of the NF-kappa B signaling pathway was examined by the dual-luciferase reporter assay system. The stable expression of c-FLIPp43 in the A375 cell lines transfected with the pCMV-Tag2B-cFLIP(p43) vector increased the protein level of NF-kappa B-p65 compared with in the A375 cell lines transfected with the empty vector. Transfection of the cells using the pCMV-Tag2B-cFLIP(p43) vector increased the amount of NF-kappa B-p65 in the nucleus in a dose-dependent manner. In conclusion, the transfection of the c-FLIPp43 expression vector induces the protein expression of NF-kappa B-p65 and promotes the activation of the NF-kappa B signaling pathway in the A375 melanoma cell line.