Nicotine treatment induces expression of matrix metalloproteinases in human osteoblastic Saos-2 cells.
Tobacco smoking is an important risk factor for the development of severe periodontitis. Recently, we showed that nicotine affected mineralized nodule formation, and that nicotine and lipopolysaccharide stimulated the formation of osteoclast-like cells by increasing production of macrophage colony-stimulating factor (M-CSF) and prostaglandin E2 (PGE2) by human osteoblastic Saos-2 cells. In the present study, we examined the effects of nicotine on the expression of matrix metalloproteinases (MMPs), tissue inhibitors of matrix metalloproteinases (TIMPs), the plasminogen activation system including the component of tissue-type plasminogen activator (tPA), urokinase-type PA (uPA), and PA inhibitor type 1 (PAI-1), alpha7 nicotine receptor, and c-fos. We also examined the effect of the nicotine antagonist D-tubocurarine on nicotine-induced expression of MMP-1. Gene expression was examined using real-time polymerase chain reaction (PCR) to estimate mRNA levels. In addition, expression of the MMP, TIMP, uPA, tPA, and PAI-1 proteins was determined by Western blotting analysis. Nicotine treatment caused expression of MMP-1, 2, 3, and 13, but not MMP-14, to increase significantly after 5 or 10 d of culture; MMP-14 expression did not change through day 14. Enhancement of MMP-1 expression by nicotine treatment was eliminated by simultaneous treatment with D-tubocurarine. In the presence of nicotine, expression of uPA, PAI-1, or TIMP-1, 2, 3, or 4 did not change over 14 d of culture, whereas expression of tPA increased significantly by day 7. Nicotine also increased expression of the alpha7 nicotine receptor and c-fos genes. These results suggest that nicotine stimulates bone matrix turnover by increasing production of tPA and MMP-1, 2, 3, and 13, thereby tipping the balance between bone matrix formation and resorption toward the latter process.
Katono
Tomoko
T
Nihon University Graduate School of Dentistry, 1-8-13, Kanda Surugadai, Tokyo 101-8310, Japan.
Kawato
Takayuki
T
Tanabe
Natsuko
N
Suzuki
Naoto
N
Yamanaka
Kazuhiro
K
Oka
Hitoshi
H
Motohashi
Masafumi
M
Maeno
Masao
M
D045744:Cell Line, Tumor; D015232:Dinoprostone / Q000378:metabolism; D005731:Ganglionic Stimulants / Q000494:pharmacology*; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000096:biosynthesis; D053509:Matrix Metalloproteinase 13 / Q000096:biosynthesis; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis; D019278:Matrix Metalloproteinase 3 / Q000096:biosynthesis; D020782:Matrix Metalloproteinases / Q000096:biosynthesis*; D009538:Nicotine / Q000494:pharmacology*; D010006:Osteoblasts / Q000201:enzymology*; D010960:Plasminogen Activators / Q000378:metabolism; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism; D010959:Tissue Plasminogen Activator / Q000096:biosynthesis
D005731:Ganglionic Stimulants; D016760:Proto-Oncogene Proteins c-fos; D009538:Nicotine; D010960:Plasminogen Activators; D010959:Tissue Plasminogen Activator; D053509:Matrix Metalloproteinase 13; D020782:Matrix Metalloproteinases; D019278:Matrix Metalloproteinase 3; D020778:Matrix Metalloproteinase 2; D020781:Matrix Metalloproteinase 1; D015232:Dinoprostone