Identification of invadopodia by TKS5 staining in human cancer lines and patient tumor samples
Identification of invadopodia by TKS5 staining in human cancer lines and patient tumor samples
复制标题
DOI：
10.1016/j.mex.2019.03.024
复制
发表时间：
2019-01-01
期刊：
影响因子：
1.9
通讯作者：
Diaz, Begona
中科院分区：
文献类型：
--
作者：
Baik, Matthew;French, Barbara;Diaz, Begona
侵袭伪足是具有蛋白水解活性的癌细胞扩张结构，为癌细胞提供重塑细胞外基质的能力。侵袭伪足促进侵袭性迁移，它们的形成与癌细胞的侵袭性和转移潜力相关。明确确定侵袭性伪足是研究侵袭性伪足调控输入、功能输出以及侵袭性伪足在癌细胞和人类肿瘤中的普遍性和意义的重要步骤。接头蛋白TKS 5是已知的侵袭伪足调节蛋白，其对于侵袭伪足的形成和活性是必需的。TKS 5在侵入伪足高度富集，并且与其他常用的侵入伪足标记物不同，它在其他类型的细胞突起中不显著积累。然而，TKS 5作为侵袭伪足的标志物的用途尚未推广，部分原因是针对人蛋白的合适抗体的可用性。我们已经评估了两种针对人TKS 5的商业抗体。在这里，我们详细的协议，用于检测侵袭足相关的TKS5在人类细胞培养和石蜡包埋存档肿瘤手术标本使用商业抗体。TKS 5染色法可用于人癌细胞系和外科手术标本中侵袭伪足的鉴定。(C)2019作者（S）由爱思唯尔公司出版
Invadopodia, cancer cell protrusive structures with associated proteolytic activity, provide cancer cells with the ability to remodel the extracellular matrix. Invadopodia facilitate invasive migration and their formation correlates with cancer cell invasiveness and metastatic potential. The unambiguous identification of invadopodia is an important step to undergo studies on invadopodia regulatory inputs, functional outputs, as well as the prevalence and significance of invadopodia for cancer cells and human tumors. The adaptor protein TKS5 is a known invadopodia regulatory protein, which is necessary for invadopodia formation and activity. TKS5 is highly enriched at invadopodia and, unlike other commonly used invadopodia markers, it does not accumulate significantly in other types of cellular protrusions. However, the use of TKS5 as a marker of invadopodia has not been generalized, in part due to the availability of suitable antibodies against the human protein. We have evaluated two commercial antibodies raised against human TKS5. Here, we detail protocols for the detection of invadopodia-associated TKS5 in human cells in culture and in paraffin-embedded archived tumor surgical specimens using commercial antibodies. These methods should facilitate the identification and study of human invadopodia.TKS5 staining identifies invadopodia in human cancer cell lines and archived surgical tumor specimens. (C) 2019 The Author(s). Published by Elsevier B.V.