The role of S100P in the invasion of pancreatic cancer cells is mediated through cytoskeletal changes and regulation of cathepsin D
The role of S100P in the invasion of pancreatic cancer cells is mediated through cytoskeletal changes and regulation of cathepsin D
复制标题
DOI：
10.1158/0008-5472.can-07-0545
复制
发表时间：
2007-09-15
期刊：
影响因子：
11.2
通讯作者：
Crnogorac-Jurcevic, Tatjana
中科院分区：
文献类型：
--
作者：
Whiteman, Hannah J.;Weeks, Mark E.;Crnogorac-Jurcevic, Tatjana
S100P是S100钙结合蛋白家族中的一员，它的上调是胰腺癌发生发展的早期分子事件，在胰腺癌前病变和浸润性癌中均有高水平表达。为了更深入地了解该蛋白的功能作用的分子机制，我们在Pancl胰腺癌细胞系中稳定过表达S100P，并通过双向凝胶差异电泳法鉴定了随后全球蛋白表达的变化。通过Western印迹分析和免疫荧光分析证实了所观察到的靶蛋白的变化，而通过运动和侵袭试验研究了它们的功能效应。在这项研究中，我们发现S100P的过表达导致了几种细胞骨架蛋白的表达水平的变化，包括细胞角蛋白8、18和19。我们还发现了肌动蛋白细胞骨架网络的紊乱和肌动蛋白调节蛋白cofilin的磷酸化状态的变化。此外，我们还发现，S100P的过度表达会导致另一种早期胰腺癌标志物S100A6和天冬氨酸蛋白酶组织蛋白D的表达增加，这两种蛋白都参与了细胞侵袭。功能研究表明，S100P过表达细胞侵袭能力的增强至少部分是由于组织蛋白酶D表达的增加。综上所述，我们的数据表明，这些变化可能有助于胰腺癌的转移扩散，并可能解释这种疾病的破坏性预后。
Up-regulation of S100P, a member of the S100 calcium-binding protein family, is an early molecular event in the development of pancreatic cancer and it is expressed at high levels in both precursor lesions and invasive cancer. To gain more insight into the molecular mechanisms underlying the functional roles of this protein, we stably overexpressed S100P in the Pancl pancreatic cancer cell line and identified the consequent changes in global protein expression by two-dimensional difference in-gel electrophoresis. The observed changes in target proteins were confirmed by Western blot analysis and immunofluorescence, whereas their functional effect was investigated using motility and invasion assays. In this study, we have shown that overexpression of S100P led to changes in the expression levels of several cytoskeletal proteins, including cytokeratins 8, 18, and 19. We have also shown disorganization of the actin cytoskeleton network and changes in the phosphorylation status of the actin regulatory protein cofilin. Additionally, we have shown that overexpression of S100P leads to increased expression of another early pancreatic cancer marker, S100A6, as well as the aspartic protease cathepsin D, both of which are involved in cellular invasion. Functional studies showed that the increased invasive potential of S100P-overexpressing cells was at least partially due to the increase in cathepsin D expression. In summary, our data suggest that these changes could contribute to the metastatic spread of pancreatic cancer and may explain the devastating prognosis of this disease.