Suppression of tumor necrosis factor alpha-induced matrix metalloproteinase 9 production by the introduction of a super-repressor form of inhibitor of nuclear factor kappaBalpha complementary DNA into immortalized human salivary gland acinar cells. Prevention of the destruction of the acinar structure in Sjögren's syndrome salivary glands.
OBJECTIVE
We have previously shown that specific enhancement in acinar cells of proteolytic activity induced by tumor necrosis factor alpha (TNFalpha) may be responsible for the destruction of the acinar structure in the salivary glands of patients with Sjögren's syndrome. Because matrix metalloproteinase 9 (MMP-9) is regulated by nuclear factor kappaB (NF-kappaB), we investigated the effect of a super-repressor form of inhibitor of nuclear factor kappaBalpha (srIkappaBalpha) on the suppression of TNFalpha-induced MMP-9 production in acinar cells.


METHODS
Two srIkappaBalpha complementary DNA (cDNA)-transfected acinar cell clones (ACMT-6 and ACMT-7) and 1 empty vector-transfected cell clone (ACpRc-1) were established. After treatment of cell clones with TNFalpha, the expression of MMP-9 was examined. In addition, the effect of TNFalpha on cell growth and the morphogenetic behavior of cell clones cultured on type IV collagen-coated dishes were examined.


RESULTS
TNFalpha induced the production of MMP-9 in the ACpRc-1 cell clone, but greatly suppressed MMP-9 production in ACMT-6 and ACMT-7 clones. No apparent cytotoxic effect of TNFalpha treatment was observed in these cell clones. When ACpRc-1 cells were seeded on type IV collagen-coated dishes in the presence of both TNFalpha and plasmin, type IV collagen interaction with the cells was lost and the cells entered apoptosis. However, even when ACMT-6 and ACMT-7 cells were cultured under the same culture conditions as those for ACpRc-1, these cell clones attached to the substrate and grew consistently without showing apoptosis. Conclusion. These observations indicate that suppression of TNFalpha-induced MMP-9 production by the introduction of srIkappaBalpha cDNA corrected the aberrant in vitro morphogenesis of acinar cells grown on type IV collagen.
Azuma
M
M
Department of Oral and Maxillofacial Surgery 2, Tokushima University School of Dentistry, Japan.
Aota
K
K
Tamatani
T
T
Motegi
K
K
Yamashita
T
T
Harada
K
K
Hayashi
Y
Y
Sato
M
M
D015153:Blotting, Western; D002455:Cell Division / Q000187:drug effects; D002999:Clone Cells / Q000737:chemistry; D003094:Collagen; D003470:Culture Media; D018076:DNA, Complementary / Q000008:administration & dosage; D005341:Fibrinolysin / Q000494:pharmacology; D015870:Gene Expression; D006801:Humans; D007150:Immunohistochemistry; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000235:genetics*; D012333:RNA, Messenger; D012469:Salivary Glands / Q000166:cytology*; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000494:pharmacology
D003470:Culture Media; D018076:DNA, Complementary; D016328:NF-kappa B; D012333:RNA, Messenger; D019715:Tissue Inhibitor of Metalloproteinase-1; D014409:Tumor Necrosis Factor-alpha; D003094:Collagen; D005341:Fibrinolysin; D020780:Matrix Metalloproteinase 9