The expression and role of non-canonical (PKC) signaling in nucleus pulposus cell metabolism
The expression and role of non-canonical (PKC) signaling in nucleus pulposus cell metabolism
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DOI：
10.1002/jor.22095
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发表时间：
2012-09-01
影响因子：
2.8
通讯作者：
Mochida, Joji
中科院分区：
文献类型：
--
作者：
Arai, Fumiyuki;Hiyama, Akihiko;Mochida, Joji
典型的Wnt/ β -连环蛋白（以下简称Wnt）信号调节多种细胞类型的增殖和分化。然而，包括蛋白激酶C （PKC）信号在内的非规范信号在椎间盘（IVD）细胞中的作用尚未被研究。本研究的目的是阐明PKC信号的激活是否能调节IVD细胞中的Wnt信号。我们使用大鼠髓核（NP）细胞进行了多种报告基因分析、实时逆转录聚合酶链反应（RT-PCR）、免疫组织化学和免疫荧光分析以及western blot分析。我们还研究了PKC信号的激活剂phorbol 12-肉豆蔻酸13-乙酸酯（PMA）刺激下的细胞增殖和细胞周期分布。我们发现NP细胞在PMA刺激下β -连环蛋白mRNA和蛋白水平下降。PMA处理以时间和剂量依赖的方式促进增殖和细胞周期进展。此外，PKC信号的激活也调节了aggrecan的表达。最后，PMA激活诱导NP细胞中几种PKC亚型的表达。由此可见，激活PKC信号可能导致基质合成和细胞增殖增加，从而抑制IVD变性。这些信号通路的串扰在IVD稳态调节中起重要作用。(C) 2012年中国骨科学会。Wiley期刊公司出版。[J]中华骨科杂志，2012,30 (4):481 - 481
Canonical Wnt/beta-catenin (hereafter Wnt) signaling regulates the proliferation and differentiation of various cell types. However, the role of non-canonical signaling including protein kinase C (PKC) signaling has not been investigated in intervertebral disc (IVD) cells. The aim of this study was to elucidate whether the activation of PKC signaling act to modulate Wnt signaling in IVD cells. We performed several reporter assays, real-time reverse transcription polymerase chain reaction (RT-PCR), immunohistochemical and immunofluorescence analyses, and western blot analyses using rat nucleus pulposus (NP) cells. We also examined the cell proliferation and cell cycle distribution under phorbol 12-myristate 13-acetate (PMA) stimulation, a known activator of PKC signaling. We found that NP cells exhibited decreased beta-catenin mRNA and protein levels upon stimulation with PMA. PMA treatment promoted proliferation and cell cycle progression in a time- and dose-dependent manner. In addition, activation of the PKC signaling also regulated the expression of aggrecan. Finally, activation by PMA induced the expression of several PKC isoforms in NP cells. It is concluded that activation of PKC signaling might lead to an increase in matrix synthesis and cell proliferation, thereby inhibiting IVD degeneration. Crosstalk in these signaling pathways plays an important role in the regulation of IVD homeostasis. (C) 2012 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 30:14781485, 2012