Upregulated expression of MMP-9 in gingival epithelial cells induced by prolonged stimulation with arecoline.
Upregulated expression of MMP-9 in gingival epithelial cells induced by prolonged stimulation with arecoline.
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DOI：
10.3892/ol.2017.6194
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发表时间：
2017-07
期刊：
影响因子：
2.9
通讯作者：
O. Uehara;Kousuke Takimoto;Tetsuro Morikawa;F. Harada;R. Takai;B. Adhikari;Ryoko Itatsu;Tomohisa Nakamura;Koki Yoshida;H. Matsuoka;H. Nagayasu;I. Saito;M. Muthumala;I. Chiba;Y. Abiko
中科院分区：
文献类型：
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作者：
O. Uehara;Kousuke Takimoto;Tetsuro Morikawa;F. Harada;R. Takai;B. Adhikari;Ryoko Itatsu;Tomohisa Nakamura;Koki Yoshida;H. Matsuoka;H. Nagayasu;I. Saito;M. Muthumala;I. Chiba;Y. Abiko
嚼槟榔液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，为了表征慢性槟榔碱刺激与癌变的关系，我们检测了基质金属蛋白酶(MMP)-2、MMP-9、组织金属蛋白酶抑制剂(TIMP)-1和TIMP-2 mRNA在槟榔碱刺激的人牙龈上皮祖细胞（HGEPs）中的表达水平。用槟榔碱（50µg/ml）孵育3天，不加槟榔碱孵育3天，最长30天。通过逆转录-定量聚合酶链反应，测定槟榔碱刺激后18天和30天细胞中MMPs和TIMPs的表达水平。与对照组相比，试验组MMP-9 mRNA表达量显著升高（P<0.01）。实验组与对照组MMP-2、TIMP-1、TIMP-2 mRNA的表达均无显著差异。通过MMP-9活性测定，实验组的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，我们使用了PDTC[核因子(NF)-κB/ NF-κB （i -κB）抑制剂的抑制剂]、PD98059[丝裂原活化蛋白激酶（MAPKK）1和MAPKK2抑制剂]、SB203580 （p38 MAPK抑制剂）和5,15- dpp[信号转导和转录激活因子（STAT） 3抑制剂]。所有抑制剂均能降低槟榔碱刺激诱导的MMP-9上调程度。基于这些数据，我们推测MMP-9的活性可能参与了咀嚼槟榔液诱导的口腔上皮病理改变，NF-κB/ i -κB、MAPK、p38 MAPK和STAT3信号通路可能参与了咀嚼槟榔液诱导的MMP-9的产生。
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.