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 kappaB elements.
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 kappaB (NFkappaB; -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. NFkappaB function was associated with specific p50- and p65-containing nuclear NFkappaB 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 NFkappaB 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.
Farina
A R
AR
Department of Experimental Medicine, University of L'Aquila, Coppito Il, Italy.
Tacconelli
A
A
Vacca
A
A
Maroder
M
M
Gulino
A
A
Mackay
A R
AR
D016227:Benzoquinones; D001665:Binding Sites; D000081247:CSK Tyrosine-Protein Kinase; D002457:Cell Extracts; D002467:Cell Nucleus / Q000378:metabolism; D017364:Collagenases / Q000235:genetics* / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells; D006801:Humans; D047029:Lactams, Macrocyclic; D020780:Matrix Metalloproteinase 9; D016328:NF-kappa B / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D009447:Neuroblastoma / Q000473:pathology*; D009474:Neurons / Q000166:cytology*; D010641:Phenotype; D011401:Promoter Regions, Genetic; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D011809:Quinones / Q000494:pharmacology; D020218:Response Elements; D017828:Rifabutin / Q000031:analogs & derivatives; D016329:Sp1 Transcription Factor / Q000378:metabolism; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014407:Tumor Cells, Cultured; D015854:Up-Regulation; D019061:src-Family Kinases
D016227:Benzoquinones; D002457:Cell Extracts; D004791:Enzyme Inhibitors; D047029:Lactams, Macrocyclic; D016328:NF-kappa B; D011809:Quinones; D016329:Sp1 Transcription Factor; D018808:Transcription Factor AP-1; D017828:Rifabutin; C020754:herbimycin; D011505:Protein-Tyrosine Kinases; D000081247:CSK Tyrosine-Protein Kinase; D019061:src-Family Kinases; C000631346:CSK protein, human; D017364:Collagenases; D020780:Matrix Metalloproteinase 9