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
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.
Oncol Lett. 2017 Jul;14(1):1186-1192. doi: 10.3892/ol.2017.6194. Epub 2017 May 17.
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.
嚼食槟榔与东南亚国家口腔癌的高发病率有关。槟榔的主要成分之一是槟榔碱。在本研究中，为了明确慢性槟榔碱刺激与致癌作用之间的关联，评估了用槟榔碱刺激的人牙龈上皮祖细胞（HGEP）中基质金属蛋白酶（MMP）-2、MMP-9、金属蛋白酶组织抑制剂（TIMP）-1和TIMP-2 mRNA的表达水平。将HGEP在含槟榔碱（50μg/ml）的培养基中培养3天与不含槟榔碱的培养基中培养3天之间交替进行，持续30天。在第18天和第30天，通过逆转录-定量聚合酶链反应评估用槟榔碱刺激的细胞中MMP和TIMP的表达水平。与对照组相比，实验组中MMP-9 mRNA的表达显著增加（P<0.01）。实验组和对照组之间在MMP-2、TIMP-1或TIMP-2 mRNA的表达上未观察到显著差异。使用MMP-9活性测定法，证明实验组中MMP-9活性水平显著高于对照组（P<0.05）。为了研究相关的细胞信号通路，使用了PDTC [一种核因子（NF）-κB/ NF-κB抑制剂（IκB）抑制剂]、PD98059 [一种丝裂原活化蛋白激酶激酶（MAPKK）1和MAPKK2抑制剂]、SB203580（一种p38丝裂原活化蛋白激酶抑制剂）和5,15-DPP [一种信号转导和转录激活因子（STAT）3抑制剂]。所有抑制剂均降低了槟榔碱刺激诱导的MMP-9上调程度。基于这些数据，推测MMP-9活性可能参与嚼食槟榔引起的口腔上皮病理改变，并且NF-κB/IκB、丝裂原活化蛋白激酶（MAPK）、p38 MAPK和信号转导和转录激活因子3（STAT3）信号通路可能参与嚼食槟榔诱导的MMP-9产生。
