사람 신경모세포종 세포주 SH-SY5Y에서 fenretinide에 의한 GD3합성효소(hST8Sia I)의 전사조절기작

Abstract

To elucidate the mechanism underlying the regulation of hST8Sia Ⅰ gene expression in FenR-induced SH-SY5Y cells, we characterized the promoter region of the hST8Sia I gene. Functional analysis of the 5’-flanking region of the hST8Sia Ⅰ gene showed that the -1146 to -646 region functions as the FenR-inducible promoter of hST8Sia Ⅰ in SH-SY5Y cells. Site-directed mutagenesis indicated that the NF-&B binding site at -731 to -722 was crucial for the FenR-induced expression of hST8Sia I in SH-SY5Y cells. To investigate which signal transduction pathway was involved in FenR-stimulated induction of hST8Sia Ⅰ in SH-SY5Y cells, we performed Western blot analysis using phospho-specific antibodies in order to measure their degree of regulatory phosphorylation. Phosphorylations of AKT and RelA (p65) subunit of NF-κB were significantly elevated in cytosolic and nuclear fractions of FenR-stimulated SH-SY5Y cells, respectively, than in control or DMSO-treated SH-SY5Y cells. These results suggest that FenR induce transcriptional up-regulation of hST8Sia I gene expression through translocation of RelA (p65) subunit of NF-κB to nucleus by AKT signal pathway in SH-SY5Y cells.