Roles of extracellular matrix tenascin-X in tumor invasion and metastasis

细胞外基质腱蛋白-X在肿瘤侵袭和转移中的作用

• 批准号: 14572048
• 负责人: MATSUMOTO Ken-ichi
• 金额: $ 2.24万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14572048/
• 关键词: Extracellular matrix; Matrix matalloproteinase; Tumor; Tenascin-X; マトリックス メタロプロテアーゼ; マトリックスメタプロテアーゼ

The results of our previous study showed that tumor invasion and metastasis are promoted in extracellular matrix tenascin-X-deficient (TNX-/-) mice via increased expression of matrix metalloproteinases (MMPs). However, little is known about the relationship between TNX deficiency and activation of MMP genes. In this study, we investigated the molecular mechanism by which TNX deficiency activates the MMP-2 gene. We examined the intracellular signaling pathways that regulate gene expression of the proteinase in isolated fibroblasts. Results of gelatin zymography showed that MMP-2 was induced to a greater extent in TNX-/-fibroblasts embedded in type I collagen than in wild-type fibroblasts RT-PCR analysis revealed that the increased level of MMP-2 expression was caused at the transcription level. Conversely, stable overexpression of TNX in a fibroblast cell line reduced MMP-2 expression and suppressed MMP-2 promoter activity. In addition, treatment of TNX-/-fibroblasts with SP600125, a c-Jun N-terminal kinase (JNK) inhibitor, and genistein, a tyrosine kinase inhibitor, suppressed the increased level of proMMP-2 and increased MMP-2 promoter activity in TNX-/-fibroblasts. Furthermore, increased activation of JNK and tyrosine phosphorylation of certain proteins were observed in TNX-/-fibroblasts. These findings suggest that induction of MMP-2 by TNX deficiency is mediated, at least in part, through the JNK and protein tyrosine kinase phosphorylation pathway.

我们以前的研究结果表明，在细胞外基质tenascin-X缺陷(TnX-/-)小鼠中，肿瘤的侵袭和转移是通过增加基质金属蛋白酶(MMPs)的表达来促进的。然而，对于Tnx缺乏与基质金属蛋白酶基因激活之间的关系却知之甚少。在本研究中，我们研究了TnX缺乏激活基质金属蛋白酶-2基因的分子机制。我们研究了在分离的成纤维细胞中调节蛋白水解酶基因表达的细胞内信号通路。明胶酶谱分析结果显示，包埋于I型胶原中的TnX-/-成纤维细胞比野生型成纤维细胞更能诱导MMP2的表达，RT-PCR分析表明MMP2的表达水平升高是在转录水平上引起的。相反，在成纤维细胞系中稳定过表达Tnx会减少基质金属蛋白酶-2的表达，并抑制基质金属蛋白酶-2的启动子活性。此外，用c-jun氨基末端激酶(JNK)抑制剂SP600125和酪氨酸激酶抑制剂金雀异黄素处理TNX-/-成纤维细胞，可抑制TnX-/-成纤维细胞中基质金属蛋白酶-2原水平的升高和基质金属蛋白酶-2启动子活性的增加。此外，在TnX-/-成纤维细胞中观察到JNK的激活和某些蛋白质的酪氨酸磷酸化增加。这些发现表明，Tnx缺乏诱导的基质金属蛋白酶-2至少部分是通过JNK和蛋白酪氨酸激酶的磷酸化途径介导的。

项目成果

Ken-ichi Matsumoto et al.: "Invasion of melanoma in double knockout mice lacking tenascin-X and tenascin-C"Jpn. J. Cancer Res.. 93巻. 968-975 (2002)

Ken-ichi Matsumoto 等：“缺乏生腱蛋白-X 和生腱蛋白-C 的双基因敲除小鼠中黑色素瘤的侵袭”Jpn. Cancer Res. 93. 968-975 (2002)

Kyoko Imanaka-Yoshida et al.: "The dynamic expression of tenascin-C and tenascin-X during early heart development in the mouse."Differentiation. 71. 291-298 (2003)

Kyoko Imanaka-Yoshida 等人：“小鼠早期心脏发育过程中腱蛋白-C 和腱蛋白-X 的动态表达。”分化。

Takeharu Minamitani et al.: "Deficiency of tenascin-X causes a decrease in the level of expression of type VI collagen."Exp.Cell Res.. (in press). (2004)

Takeharu Minamitani 等人：“腱蛋白-X 的缺乏会导致 VI 型胶原蛋白表达水平下降。”Exp.Cell Res..（出版中）。

Ken-ichi Matsumoto et al.: "Distribution of extracellular matrix tenascin-X in sciatic nerves."Acta Neuropathol.. 104. 448-454 (2002)

Ken-ichi Matsumoto 等人：“坐骨神经中细胞外基质腱蛋白-X 的分布。”Acta Neuropathol.. 104. 448-454 (2002)

Ken-ichi Matsumoto et al.: "Induction of matrix metalloproteinase-2 by tenascin-X deficiency is mediated through the c-Jun N-terminal kinase and protein tyrosine kinase phosphorylation pathway."Exp.Cell.Res.. (in press). (2004)

Ken-ichi Matsumoto 等人：“腱蛋白-X 缺陷对基质金属蛋白酶-2 的诱导是通过 c-Jun N 末端激酶和蛋白酪氨酸激酶磷酸化途径介导的。”Exp.Cell.Res..（出版中）