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Transcriptional Up-Regulation of Matrix Metalloproteinase-9 Expression during Spontaneous Epithelial to Neuroblast Phenotype Conversion by SK-N-SH Neuroblastoma Cells, Involved in Enhanced Invasivity, Depends upon GT-Box and Nuclear Factor B Elements¹

Laboratory of Molecular Pathology, Department of Experimental Medicine, University of L’Aquila, Coppito II, 67100 L’Aquila [A. R. F., A. T., A. R. M.]; Neuromed Institute, 80077 Pozzilli [A. R. F., A. G., A. R. M.]; Department of Experimental Medicine and Pathology, University of Rome "La Sapienza," 00161 Rome [A. V., A. G.]; and Institute of Pathology, University of Palermo, 90141 Palermo [M. M.], Italy

Spontaneous epithelial (S) to neuroblast (N) conversion enhanced the capacity of SK-N-SH neuroblastoma (NB) cells to invade reconstituted basement membrane in vitro. This involved a switch to matrix metalloproteinase (MMP) activity, in particular MMP-9, and was associated with the induction of MMP-9 expression. N-type-specific MMP-9 expression was herbimycin A inhibitable tyrosine kinase (possibly c-src) dependent and was regulated transcriptionally through GT-box (-52), and nuclear factor B (NFB; -600) elements within the MMP-9 gene. GT-box function was associated with elevated levels of specific nuclear GT-box binding complexes in N-type cells. NFB function was associated with specific p50- and p65-containing nuclear NFB binding complex(es). No function could be attributed to the proximal AP-1 (-79) element, and minimal function was attributed to the SP-1 (-560), ets (-540), or distal AP-1 (-533) elements. This was despite elevated levels of specific junD/fra-1 containing proximal AP-1 element binding complex(es) in N-type cells. Our data highlight a pivotal role for the GT-box, in concert with the NFB element, in the transcriptional up-regulation of MMP-9 expression during spontaneous S to N phenotype conversion by SK-N-SH cells involved in enhanced basement membrane invasivity.

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