Induction of collagenase-2 (matrix metalloproteinase-8) gene expression by interleukin-1beta in human gingival fibroblasts.
Collagenase-2 (matrix metalloproteinase-8 or MMP-8) is synthesized mainly by polymorphonuclear neutrophils and plays a crucial role in inflammatory periodontal tissue destruction. We tested the effect of interleukin(IL)-1beta, a proinflammatory cytokine, on collagenase-2 gene expression in cultured human gingival fibroblasts and also compared this effect with IL-1beta-induced changes in collagenase-1 and -3 gene expression. By a combination of reverse transcription-polymerase chain reaction and Southern analysis, IL-1beta was found to dose-dependently induce gene expression for collagenase-1, -2, and -3 in gingival fibroblasts. Although collagenase-2 mRNA was the least abundant among the three collagenase mRNAs tested in the cultured fibroblast system, addition of 1 ng/ml IL-1beta significantly increased collagenase-2 gene transcription within 6 h, and maximal stimulation was maintained for 12 to 48 h. Significant mRNA induction was observed with as little as 0.1 ng/ml IL-1beta. IL-1beta was also found to increase the stability of collagenase-2 mRNAs after transcription arrest was induced by an RNA polymerase inhibitor. Stimulation of collagenase-2 mRNA expression by IL-1beta was prevented by pretreatment with cycloheximide, an inhibitor of protein synthesis. These results indicate that IL-1beta increased mRNA expression for collagenases including collagenase-2 in gingival fibroblasts. The findings also suggest that enhancement of collagenase-2 mRNA expression by IL-1beta involves both protein synthesis and suppression of mRNA degradation.
Abe
M
M
Department of Biochemistry, Ohu University School of Dentistry, Koriyama, Japan.
Kawamoto
K
K
Okamoto
H
H
Horiuchi
N
N
D000704:Analysis of Variance; D015139:Blotting, Southern; D018929:Cell Culture Techniques; D017364:Collagenases / Q000235:genetics; D003513:Cycloheximide / Q000494:pharmacology; D012321:DNA-Directed RNA Polymerases / Q000037:antagonists & inhibitors; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000201:enzymology*; D015971:Gene Expression Regulation, Enzymologic; D005881:Gingiva / Q000201:enzymology*; D006801:Humans; D018836:Inflammation Mediators / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology*; D053509:Matrix Metalloproteinase 13; D020784:Matrix Metalloproteinase 8 / Q000235:genetics*; D009504:Neutrophils / Q000201:enzymology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013223:Statistics as Topic; D014158:Transcription, Genetic
D018836:Inflammation Mediators; D007375:Interleukin-1; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D003513:Cycloheximide; D012321:DNA-Directed RNA Polymerases; D017364:Collagenases; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13; C417249:collagenase 1; D020784:Matrix Metalloproteinase 8