Upregulated expression of MMP-9 in gingival epithelial cells induced by prolonged stimulation with arecoline

Uehara,Osamu; Takimoto,Kousuke; Morikawa,Tetsuro; Harada,Fumiya; Takai,Rie; Adhikari,Bhoj  Raj; Itatsu,Ryoko; Nakamura,Tomohisa; Yoshida,Koki; Matsuoka,Hirofumi; Nagayasu,Hiroki; Saito,Ichiro; Muthumala,Malsantha; Chiba,Itsuo; Abiko,Yoshihiro

doi:10.3892/ol.2017.6194

Upregulated expression of MMP-9 in gingival epithelial cells induced by prolonged stimulation with arecoline

Authors:
- Osamu Uehara
- Kousuke Takimoto
- Tetsuro Morikawa
- Fumiya Harada
- Rie Takai
- Bhoj Raj Adhikari
- Ryoko Itatsu
- Tomohisa Nakamura
- Koki Yoshida
- Hirofumi Matsuoka
- Hiroki Nagayasu
- Ichiro Saito
- Malsantha Muthumala
- Itsuo Chiba
- Yoshihiro Abiko
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Affiliations: Division of Disease Control and Molecular Epidemiology, Department of Oral Growth and Development, School of Dentistry, Health Sciences University of Hokkaido, Tobetsu, Hokkaido 061‑0293, Japan, Division of Oral and Maxillofacial Surgery, Department of Human Biology and Pathophysiology, School of Dentistry, Health Sciences University of Hokkaido, Tobetsu, Hokkaido 061‑0293, Japan, Division of Oral Medicine and Pathology, Department of Human Biology and Pathophysiology, School of Dentistry, Health Sciences University of Hokkaido, Tobetsu, Hokkaido 061‑0293, Japan, Department of Pathology, Tsurumi University School of Dental Medicine, Yokohama, Kanagawa 230‑8501, Japan, Department of Oral and Maxillofacial Surgery, Army Hospital, Colombo, Sri Lanka
Published online on: May 17, 2017 https://doi.org/10.3892/ol.2017.6194
Pages: 1186-1192

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Abstract

Betel quid chewing is implicated in the high prevalence of oral cancer in Southeast Asian countries. One of the major components of betel quid is arecoline. In the present study, in order to characterize the association between chronic arecoline stimulation and carcinogenesis the expression level of matrix metalloproteinase (MMP)‑2, MMP‑9, tissue inhibitor of metalloproteinase (TIMP)‑1 and TIMP‑2 mRNA in human gingival epithelial progenitor cells (HGEPs) stimulated with arecoline was assessed. The HGEPs were alternated between 3 days of incubation with arecoline (50 µg/ml), and 3 days without arecoline, for up to 30 days. The expression levels of the MMPs and TIMPs in the cells stimulated with arecoline were evaluated by reverse transcription-quantitative polymerase chain reaction at 18 and 30 days. The expression of MMP‑9 mRNA in the experimental group was significantly increased compared with in the control group (P<0.01). No significant differences in the expression of MMP‑2, TIMP‑1 or TIMP‑2 mRNA were observed between the experimental and control groups. Using an MMP‑9 activity assay, the levels of MMP‑9 activity in the experimental group were demonstrated to be significantly higher than in the control group (P<0.05). To investigate associated cellular signaling pathways, PDTC [a nuclear factor (NF)‑κB/inhibitor of NF‑κB (IκB) inhibitor], PD98059 [a mitogen-activated protein kinase kinase (MAPKK)1 and MAPKK2 inhibitor], SB203580 (a p38 MAPK inhibitor) and 5,15‑DPP [a signal transduction and activator of transcription (STAT) 3 inhibitor] were used. All inhibitors decreased the extent of MMP‑9 upregulation induced by stimulation with arecoline. Based on the data, it is hypothesized that MMP‑9 activity may be involved in the pathological alterations of oral epithelium induced by betel quid chewing, and that the NF‑κB/IκB, MAPK, p38 MAPK and STAT3 signaling pathways may be involved in the production of MMP-9 induced by betel quid chewing.

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