The role of lamin A in cytoskeleton organization in colorectal cancer cells A proteomic investigation
The role of lamin A in cytoskeleton organization in colorectal cancer cells A proteomic investigation
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DOI：
10.4161/nucl.2.5.17775
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发表时间：
2011-09-01
期刊：
影响因子：
3.7
通讯作者：
Hutchison, Christopher J.
中科院分区：
文献类型：
--
作者：
Foster, Clare R.;Robson, Joanne L.;Hutchison, Christopher J.
核纤层蛋白A的上调表达与结直肠癌中细胞侵袭性和死亡率的增加有关。在这里，我们使用定量蛋白质组学研究核纤层蛋白A调节的细胞骨架的变化，可能支持增加细胞运动。在模型细胞系（SW 480/lamA）中使用核纤层蛋白A的siRNA敲低，我们证实核纤层蛋白A的存在促进细胞运动。使用增强的技术来制备细胞骨架组分结合2D DiGE，我们能够准确和可重复地检测低至20%的细胞骨架内蛋白质种类的代表性变化。与对照组相比，SW 480/lamA细胞的细胞骨架中共有64个蛋白质点显示出增加或减少的表达。其中29个斑点的身份通过质谱法确定。大多数是三类蛋白质的多种形式，包括肌动蛋白和IF细胞骨架的组分、蛋白伴侣和翻译起始和延伸因子。特别地，我们的数据揭示了组织转氨酶2的表达，其已知修饰细胞骨架的元件并且与癌症进展相关，在SW 480/lamA细胞的细胞骨架部分中高度过度表达。总的来说，我们的数据是一致的改变蛋白质交联和折叠，有利于形成动态肌动蛋白丝应力纤维占改变细胞运动特性在SW 480/lamA细胞。
Upregulated expression of lamin A has been implicated in increased cell invasiveness and mortality in colorectal cancer. Here we use quantitative proteomics to investigate lamin A regulated changes in the cytoskeleton that might underpin increased cell motility. Using siRNA knockdown of lamin A in a model cell line (SW480/lamA) we confirm that the presence of lamin A promotes cell motility. Using an enhanced technique to prepare cytoskeleton fractions in combination with 2D DiGE we were able to accurately and reproducibly detect changes in the representation of protein species within the cytoskeleton as low as 20%. In total 64 protein spots displayed either increased or decreased representation within the cytoskeleton of SW480/lamA cells compared to controls. Of these the identities of 29 spots were determined by mass spectrometry. A majority were multiple forms of three classes of proteins, including components of the actin and IF cytoskeletons, protein chaperones and translation initiation and elongation factors. In particular our data reveal that the representation of tissue transglutaminase 2, which is known to modify elements of the cytoskeleton and is associated with cancer progression, was highly over-represented in the cytoskeleton fraction of SW480/lamA cells. Overall, our data are consistent with changed protein cross-linking and folding that favours the formation of dynamic actin filaments over stress fibers accounting for the altered cell motility properties in SW480/lamA cells.