Enhancement of intervertebral disc cell senescence by WNT/β-catenin signaling-induced matrix metalloproteinase expression.
Enhancement of intervertebral disc cell senescence by WNT/β-catenin signaling-induced matrix metalloproteinase expression.
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DOI：
10.1002/art.27599
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发表时间：
2010-10
影响因子：
--
通讯作者：
Mochida, Joji
中科院分区：
文献类型：
--
作者：
Hiyama, Akihiko;Sakai, Daisuke;Risbud, Makarand V.;Tanaka, Masahiro;Arai, Fumiyuki;Abe, Koichiro;Mochida, Joji
目的：研究椎间盘细胞是否表达β-连环蛋白，并探讨β-连环蛋白信号通路在细胞衰老和聚集素合成中的作用。采用实时定量逆转录聚合酶链式反应、免疫印迹、免疫组织化学和免疫荧光等方法检测β-catenin信使核糖核酸和蛋白在大鼠脑内血管内皮细胞中的表达。通过转染实验、四甲基偶氮唑盐比色法、衰老相关β半乳糖苷酶染色、细胞周期分析和转化生长因子/骨形态发生蛋白(ββ)/骨形态发生蛋白(BMP)通路芯片分析，检测WNT/BMP-catenin对髓核(NP)细胞的影响。我们发现β-catenin基因和蛋白在体内的视盘中表达，并且大鼠NP细胞在氯化锂刺激下β-catenin基因和蛋白表达增加。LiCl处理对NP细胞的增殖有抑制作用，呈时间和剂量依赖性。此外，氯化锂处理的细胞衰老程度增加。长期应用LiCl2诱导NP细胞周期停滞，促进随后的细胞凋亡。Wnt/β-catenin信号的激活也调节了aggrecan的表达。我们还证实了Wnt/β-catenin信号诱导了NP细胞中基质金属蛋白酶和转化生长因子β的表达。Wnt/β-catenin信号通路的激活促进了细胞的衰老，可能调节了NP细胞中基质金属蛋白酶和转化生长因子β信号的表达。我们推测，Wnt/β-catenin信号的激活可能导致基质的分解增加，从而促进血管性痴呆的退变。
To determine whether intervertebral disc (IVD) cells express β-catenin and to assess the role of the WNT/β-catenin signaling pathway in cellular senescence and aggrecan synthesis. The expression of β-catenin messenger RNA (mRNA) and protein in rat IVD cells was assessed by using several real-time reverse transcription–polymerase chain reaction, Western blot, immunohistochemical, and immunofluorescence analyses. The effect of WNT/β-catenin on nucleus pulposus (NP) cells was examined by transfection experiments, an MTT assay, senescence-associated β-galactosidase staining, a cell cycle analysis, and a transforming growth factor (TGFβ)/bone morphogenetic protein (BMP) pathway–focused microarray analysis. We found that β-catenin mRNA and protein were expressed in discs in vivo and that rat NP cells exhibited increased β-catenin mRNA and protein upon stimulation with lithium chloride, a known activator of WNT signaling. LiCl treatment inhibited the proliferation of NP cells in a time- and dose-dependent manner. In addition, there was an increased level of cellular senescence in LiCl-treated cells. Long-term treatment with LiCl induced cell cycle arrest and promoted subsequent apoptosis in NP cells. Activation of WNT/β-catenin signaling also regulated the expression of aggrecan. We also demonstrated that WNT/β-catenin signaling induced the expression of matrix metalloproteinases (MMPs) and TGFβ in NP cells. The activation of WNT/β-catenin signaling promotes cellular senescence and may modulate MMP and TGFβ signaling in NP cells. We hypothesize that the activation of WNT/β-catenin signaling may lead to an increased breakdown of the matrix, thereby promoting IVD degeneration.
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影响因子：
3.7
作者：
HAYFLICK, L;MOORHEAD, PS
通讯作者：
MOORHEAD, PS
影响因子：
3
作者：
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作者：
Hiyama, Akihiko;Mochida, Joji;Sakai, Daisuke
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Sakai, Daisuke
影响因子：
15.9
作者：
Ben-Porath, I;Weinberg, RA
通讯作者：
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