Increased matrix metalloproteinase-2 activity induced by TGF-beta 1 in duct cells of human salivary gland is associated with the development of cyst formation in vivo.
Proteolytic enzyme activity has been shown to be important for cyst formation. In this study, we constructed a cyst-like structure in vivo and analyzed molecular mechanisms involved in the development of the lesion. When SV40-immortalized duct cells of normal human salivary gland (NS-SV-DC) were treated with TGF-beta 1 at a concentration of 1 ng/ml or 5 ng/ml followed by co-inoculation with Matrigel into the backs of nude mice, they formed large cysts containing fluid when 5 ng/ml of TGF-beta 1 was used. Analysis of the fluid demonstrated high MMP activity. Immunohistochemical staining exhibited strong reactivity with anti-MMP-2 antibody in TGF-beta 1 (5 ng/ml)-treated NS-SV-DC. Northern blot analysis indicated that the expression of TGF-beta 1 and MMP-2 mRNAs in cells was greatly enhanced by treatment with 5 ng/ml TGF-beta 1. These findings suggest that the in vivo cyst formation by TGF-beta 1-treated cells is associated with continuous induction of MMP-2 activity.
Azuma
M
M
Second Department of Orai and Maxillofacial Surgery, Tokushima University School of Dentistry, Japan.
Tamatani
T
T
Yuki
T
T
Motegl
K
K
Hoque
M O
MO
Yoshida
H
H
Sato
M
M
D000818:Animals; D015152:Blotting, Northern; D002460:Cell Line; D003094:Collagen; D004396:Coloring Agents; D003560:Cysts / Q000201:enzymology / Q000209:etiology*; D004338:Drug Combinations; D005109:Extracellular Matrix; D005260:Female; D018093:Gelatinases / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D015971:Gene Expression Regulation, Enzymologic; D006023:Glycoproteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007797:Laminin; D020778:Matrix Metalloproteinase 2; D061965:Matrix Metalloproteinase Inhibitors; D008666:Metalloendopeptidases / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008967:Molecular Biology; D011480:Protease Inhibitors / Q000032:analysis / Q000378:metabolism; D011509:Proteoglycans; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D018987:Salivary Ducts / Q000201:enzymology* / Q000473:pathology; D012466:Salivary Gland Diseases / Q000201:enzymology / Q000209:etiology; D019714:Tissue Inhibitor of Metalloproteinases; D016212:Transforming Growth Factor beta / Q000032:analysis / Q000235:genetics / Q000494:pharmacology*
D004396:Coloring Agents; D004338:Drug Combinations; D006023:Glycoproteins; D007797:Laminin; D061965:Matrix Metalloproteinase Inhibitors; D011480:Protease Inhibitors; D011509:Proteoglycans; D012333:RNA, Messenger; D019714:Tissue Inhibitor of Metalloproteinases; D016212:Transforming Growth Factor beta; C059485:matrigel; D003094:Collagen; D018093:Gelatinases; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2