中性粒细胞外陷阱在癌细胞粘附中的动态作用和Notch 1介导的上皮到间质转变的激活EGFR驱动的肺癌细胞

中性粒细胞外陷阱（NET）是激活的中性粒细胞释放的复杂结构，可能会调节转移性级联反应的不同步骤。我们研究的目的是研究网络如何调节癌细胞的粘附特性，以及细胞暴露于网络可以激活上皮到间质转变（EMT）程序，从而增强肿瘤细胞的迁移和侵入性特性。使用固定元素或不使用Net codib的固定元素，不适合使用固定元素或不使用固定元素，不适合使用固定元素或不使用固定剂量。 CCDC25受体。孵育1-4小时后，粘附细胞表示为总细胞数的百分比。为了测试EMT的发生，将细胞用网络处理长达48小时，然后用E-钙粘蛋白，波形蛋白，蜗牛，sl，slug，Zeb 1和Twist 1以及Notch 1和Notch 1的水平进行处理。使用带有Transwell和FBS作为化学吸引剂的24个Multiwell板对未经处理和网络处理的细胞进行迁移测定。癌细胞粘附在净涂层板上的粘附在30％至92.7％之间，并且显着高于未涂层板中获得的净涂层。添加针对α5β1或CCDC25的抗体导致细胞粘附对网。 EGFR驱动的癌细胞系对网的长时间暴露于NET引起了EMT程序的激活，通过上调和裂解Notch 1，并通过EMT标记的增强表达证实。随之而来的上皮表型的丧失导致癌基因驱动器的表达大大降低。与未经处理的细胞相比，网络处理细胞的细胞迁移显着增强。它的发现揭示了网络的动态作用，该网络的动态作用可能提供了富于DNA和纤连蛋白的环境，从而在远处的位点结合了许多癌细胞，在远距离接触的网站上，延长到网络上的网络暴露于NETS通过Notch 1信号通道的激活触发EMT，并具有随后的癌症和癌症的迁移性。此外，我们的发现提供了一个例子，说明了免疫/炎症微环境如何直接调节癌细胞对癌细胞靶向剂的敏感性。

Neutrophil extracellular traps (NETs) are complex structures released by activated neutrophils that may modulate different steps of the metastatic cascade. The aim of our study was to investigate how NETs can modulate the adhesion properties of cancer cells and whether cell exposure to NETs can activate the epithelial-to-mesenchymal transition (EMT) program thus enhancing the migratory and invasive properties of tumor cells.Different cancer cell lines were subjected to a solid-phase adhesion assay using NET-coated plates with or without the addition of antibodies against α5β1 or CCDC25 receptor. After 1-4 h of incubation, adherent cells were expressed as the percentage of total cell number. To test EMT occurrence, cells were treated with NETs for up to 48 h and then the levels of E-cadherin, vimentin, Snail, Slug, Zeb 1 and Twist 1 along with levels of Notch 1 and cleaved Notch 1 were determined by western blotting. Untreated and NET-treated cells were subjected to migration assays using 24-multiwell plates with transwell and FBS as chemoattractant.Cancer cell adhesion to NET-coated plates varied between 30% and 92.7% and was significantly higher than that obtained in uncoated plates. The addition of antibodies against α5β1 or CCDC25 caused a strong reduction of cell adhesion to NETs. The prolonged exposure of EGFR-driven cancer cell lines to NETs caused the activation of the EMT program through the upregulation and cleavage of Notch 1 and was confirmed by the enhanced expression of EMT markers. The consequent loss of the epithelial phenotype induced a strong reduction of the expression of the oncogene driver. Cell migration was significantly enhanced in NET-treated cells as compared to untreated cells.Our findings reveal the dynamic role of NETs that may provide a DNA and fibronectin rich environment for binding of many cancer cells at distant sites where the prolonged exposure to NETs triggers the EMT through the activation of Notch 1 signaling pathway with the subsequent enhancement of migratory and invasive properties of cancer cells. Furthermore, our findings provide an example of how an immune/inflammatory microenvironment may directly modulate the sensitivity of cancer cells to oncogene targeted agents.