Quantitative analysis of keratin filament networks in scanning electron microscopy images of cancer cells
Quantitative analysis of keratin filament networks in scanning electron microscopy images of cancer cells
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DOI：
10.1111/j.1365-2818.2005.01505.x
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发表时间：
2005-11-01
影响因子：
2
通讯作者：
Walther, P
中科院分区：
文献类型：
--
作者：
Beil, M;Braxmeier, H;Walther, P
角蛋白丝网络是细胞骨架的重要组成部分。它参与调节上皮细胞的形状和粘弹性。角蛋白网络的形态取决于角蛋白单体的翻译后修饰。体外研究表明，网络特征，如长丝交联密度，决定了长丝网络的生物物理性质。本文提出了一种定量分析角蛋白纤维网络形态的方法。细丝的可视化是基于上皮细胞的预固定提取和扫描电子显微镜（SEM）。SEM图像通过卷积算法处理，以获得表示单个细丝及其连接的图形结构。本研究应用该方法研究转化生长因子α（TGF α）对胰腺癌细胞角蛋白网络形态的影响。TGF α促进胰腺癌进展并激活磷酸化角蛋白单体的信号传导途径。使用这种新方法，可以检测到响应于TGF α的角蛋白网络形态的显著改变。
The keratin filament network is an important part of the cytoskeleton. It is involved in the regulation of shape and viscoelasticity of epithelial cells. The morphology of keratin networks depends on post-translational modifications of keratin monomers. In-vitro studies indicated that network characteristics, such as filament crosslink density, determines the biophysical properties of the filament network. This report presents a quantitative method for the morphological analysis of keratin filament networks. Visualization of filaments was based on prefixation extraction of epithelial cells and scanning electron microscopy (SEM). SEM images were processed by a skeletonization algorithm to obtain a graph structure that represents individual filaments as well as their connections. This method was applied to investigate the effects of transforming growth factor alpha (TGF alpha) on the morphology of keratin networks in pancreatic cancer cells. TGF alpha contributes to pancreatic cancer progression and activates signalling pathways phosphorylating keratin monomers. Using this new method, a significant alteration to the keratin network morphology could be detected in response to TGF alpha.