c-[FLIP.sub.p43] induces activation of the nuclear factor-κB signaling pathway in a dose-dependent manner in the A375 melanoma cell line

• Published in: Molecular medicine reports Vol. 10; no. 3; p. 1438
• Main Authors: Wang, Yujue, Tu, Yating, Lu, Jiejie, Tao, Juan, Li, Yan
• Format: Journal Article
• Language: English
• Published: Spandidos Publications 01-09-2014
• Subjects: Genetic aspects; Melanoma; Transcription factors; China
• ISSN: 1791-2997
• DOI: 10.3892/mmr.2014.2364

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