Matrix metalloproteinase-3 in odontoblastic cells derived from ips cells: unique proliferation response as odontoblastic cells derived from ES cells.
Matrix metalloproteinase-3 in odontoblastic cells derived from ips cells: unique proliferation response as odontoblastic cells derived from ES cells.
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DOI：
10.1371/journal.pone.0083563
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Nakamura H
中科院分区：
文献类型：
--
作者：
Hiyama T;Ozeki N;Mogi M;Yamaguchi H;Kawai R;Nakata K;Kondo A;Nakamura H
我们之前报道过基质金属蛋白酶 (MMP)-3 可加速牙髓损伤后的伤口愈合。此外，我们报道了促炎细胞因子混合物（肿瘤坏死因子-α、白细胞介素（IL）-1β和干扰素-γ）诱导小鼠胚胎干（ES）细胞来源的成牙本质细胞样细胞中的MMP-3活性，表明MMP-3在成牙本质细胞样细胞的伤口愈合和牙髓再生中发挥潜在的独特生理作用。在本研究中，我们测试了这样的假设：IL-1β 上调 MMP-3 活性可促进源自诱导多能干 (iPS) 和 ES 细胞的纯化成牙本质细胞样细胞的增殖和凋亡。分离每个成牙本质细胞样细胞并与不同浓度的 IL-1β 一起孵育。分别使用 RT-PCR 和蛋白质印迹评估 MMP-3 mRNA 和蛋白表达。使用免疫沉淀和荧光底物测量 MMP-3 活性。分别使用 ELISA 检测 BrdU 和 DNA 片段来测定细胞增殖和凋亡。 siRNA 用于减少这些细胞中的 MMP-3 转录物。 IL-1β 治疗增加了成牙本质细胞样细胞中 MMP-3 mRNA 和蛋白水平以及 MMP-3 活性。发现细胞增殖显着增加，而细胞凋亡没有变化。在所有实验中，内源性组织金属蛋白酶抑制剂 (TIMP)-1 和 TIMP-2 均持续表达。胞吐作用抑制剂 Exo1 有效抑制条件培养基中 MMP-3 的出现。用针对MMP-3的siRNA治疗抑制了IL-1β诱导的MMP-3表达和活性的增加，并且还抑制了细胞增殖，但意外地增加了这些细胞的凋亡（P<0.05）。研究发现外源性 MMP-3 可诱导源自 iPS 细胞和 ES 细胞的成牙本质细胞样细胞的细胞增殖。这种 siRNA 介导的细胞凋亡增加可以通过外源 MMP-3 刺激来逆转 (P<0.05)。总而言之，IL-1β 诱导 MMP-3 调节的细胞增殖并抑制源自 iPS 和 ES 细胞的成牙本质细胞样细胞的凋亡。
We previously reported that matrix metalloproteinase (MMP)-3 accelerates wound healing following dental pulp injury. In addition, we reported that a proinflammatory cytokine mixture (tumor necrosis factor-α, interleukin (IL)-1β and interferon-γ) induced MMP-3 activity in odontoblast-like cells derived from mouse embryonic stem (ES) cells, suggesting that MMP-3 plays a potential unique physiological role in wound healing and regeneration of dental pulp in odontoblast-like cells. In this study, we tested the hypothesis that upregulation of MMP-3 activity by IL-1β promotes proliferation and apoptosis of purified odontoblast-like cells derived from induced pluripotent stem (iPS) and ES cells. Each odontoblast-like cell was isolated and incubated with different concentrations of IL-1β. MMP-3 mRNA and protein expression were assessed using RT-PCR and western blotting, respectively. MMP-3 activity was measured using immunoprecipitation and a fluorescence substrate. Cell proliferation and apoptosis were determined using ELISA for BrdU and DNA fragmentation, respectively. siRNA was used to reduce MMP-3 transcripts in these cells. Treatment with IL-1β increased MMP-3 mRNA and protein levels, and MMP-3 activity in odontoblast-like cells. Cell proliferation was found to markedly increase with no changes in apoptosis. Endogenous tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2 were constitutively expressed during all experiments. The exocytosis inhibitor, Exo1, potently suppressed the appearance of MMP-3 in the conditioned medium. Treatment with siRNA against MMP-3 suppressed an IL-1β-induced increase in MMP-3 expression and activity, and also suppressed cell proliferation, but unexpectedly increased apoptosis in these cells (P<0.05). Exogenous MMP-3 was found to induce cell proliferation in odontoblast-like cells derived from iPS cells and ES cells. This siRNA-mediated increase in apoptosis could be reversed with exogenous MMP-3 stimulation (P<0.05). Taken together, IL-1β induced MMP-3-regulated cell proliferation and suppressed apoptosis in odontoblast-like cells derived from iPS and ES cells.
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影响因子：
7.6
作者：
Goda, S.;Inoue, H.;Domae, N.
通讯作者：
Domae, N.
影响因子：
3.7
作者：
Eba H;Murasawa Y;Iohara K;Isogai Z;Nakamura H;Nakamura H;Nakashima M
通讯作者：
Nakashima M
影响因子：
3
作者：
Ozeki, N;Mogi, M;Togari, A
通讯作者：
Togari, A
DOI：
10.1073/pnas.1204991109
发表时间：
2012-07-31
影响因子：
11.1
作者：
Manka, Szymon W.;Carafoli, Federico;Nagase, Hideaki
通讯作者：
Nagase, Hideaki
影响因子：
4.8
作者：
Mogi, M;Togari, A
通讯作者：
Togari, A