All-trans-retinoic acid induces nuclear factor kappaB activation and matrix metalloproteinase-9 expression and enhances basement membrane invasivity of differentiation-resistant human SK-N-BE 9N neuroblastoma Cells.
A comparison between retinoic acid (RA) differentiation-resistant and differentiation-sensitive SK-N-BE neuroblastoma (NB) cell lines revealed an association between resistance to differentiation, exhibited by N-myc stable transfected SK-N-BE 9N cells, with sensitivity to RA induction of p50/p65 nuclear factor kappaB (NF-kappaB) transcription factor activity and induction of matrix metalloproteinase (MMP)-9 expression leading to enhanced invasive behavior in vitro. These effects were not observed in differentiation-sensitive parental SK-N-BE or control-transfected SK-N-BE 2N counterparts. RA activated a MMP-9 promoter reporter gene construct in SK-N-BE 9N but not parental SK-N-BE or SK-N-BE 2N cells through a NF-kappaB element (-600) in association with enhanced p50 mRNA expression, reduced cytoplasmic inhibitor of nuclear factor kappaBalpha protein levels, and the induction of nuclear p50/p65 containing MMP-9 NF-kappaB site binding activity. RA activation of the MMP-9 promoter was inhibited by transient overexpression of a dominant-negative inhibitor of nuclear factor kappaBalpha protein and stimulated by transient p50 but not p65 overexpression in the absence of RA. A limited, nonessential function for activator protein 1 (-74), Ets (-540), and SP1 (-560) elements within the MMP-9 promoter was revealed by point mutation but was not associated with changes in the binding or position of complexes constitutive to differentiation-sensitive or -resistant cells. Our data indicates that in this model of NB resistance to differentiation that results from uncoupled RA regulation of N-myc expression, RA stimulates malignant NB cell behavior by inducing nuclear NF-kappaB transcription factor activity, which in turn induces MMP-9 expression and stimulation of basement membrane invasive capacity involving MMP-9 activity.
Section of Molecular Pathology, Department of Experimental Medicine, University of L'Aquila, 67100 L'Aquila, Italy.
Antonietta R
AR
Farina
Maria-Paola
MP
Masciulli
Antonella
A
Tacconelli
Lucia
L
Cappabianca
Giuseppina
G
De Santis
Alberto
A
Gulino
Andrew R
AR
Mackay
D000970:Antineoplastic Agents / Q000494:pharmacology*; D001485:Basement Membrane / Q000187:drug effects / Q000378:metabolism*; D001672:Biocompatible Materials; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002648:Child; D003094:Collagen; D004338:Drug Combinations; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000502:physiology; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D007797:Laminin; D020780:Matrix Metalloproteinase 9 / Q000235:genetics*; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000235:genetics* / Q000378:metabolism*; D052002:NF-kappa B p50 Subunit; D009361:Neoplasm Invasiveness / Q000235:genetics*; D009447:Neuroblastoma / Q000201:enzymology / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics; D011509:Proteoglycans; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D001672:Biocompatible Materials; D004338:Drug Combinations; D020672:I-kappa B Proteins; D007797:Laminin; D016328:NF-kappa B; D052002:NF-kappa B p50 Subunit; C000606204:NFKBIA protein, human; D011509:Proteoglycans; D012333:RNA, Messenger; C059485:matrigel; D000072000:NF-KappaB Inhibitor alpha; D014212:Tretinoin; D003094:Collagen; D020780:Matrix Metalloproteinase 9