Schelch Karin, Vogel Lisa, Schneller Anja, Brankovic Jelena, Mohr Thomas, Mayer Rupert L, Slany Astrid, Gerner Christopher, Grusch Michael
Institute of Cancer Research, Department of Medicine I, Medical University of Vienna, Vienna, Austria.
Department of Analytical Chemistry, Faculty of Chemistry, University of Vienna, Vienna, Austria.
Front Cell Dev Biol. 2021 Mar 12;9:634371. doi: 10.3389/fcell.2021.634371. eCollection 2021.
Tumors and the tumor microenvironment produce multiple growth factors that influence cancer cell behavior via various signal transduction pathways. Growth factors, like transforming growth factor β (TGFβ) and epidermal growth factor (EGF), have been shown to induce proliferation, migration, and invasion in different cell models. Both factors are frequently overexpressed in cancer and will often act in combination. Although both factors are being used as rational targets in clinical oncology, the similarities and differences of their contributions to cancer cell migration and invasion are not fully understood. Here we compared the impact of treating A549 lung adenocarcinoma cells with TGFβ, EGF, and both in combination by applying videomicroscopy, functional assays, immunoblotting, real-time PCR, and proteomics. Treatment with both factors stimulated A549 migration to a similar extent, but with different kinetics. The combination had an additive effect. EGF-induced migration depended on activation of the mitogen-activated protein kinase (MAPK) pathway. However, this pathway was dispensable for TGFβ-induced migration, despite a strong activation of this pathway by TGFβ. Proteome analysis (data are available via ProteomeXchange with identifier PXD023024) revealed an overlap in expression patterns of migration-related proteins and associated gene ontology (GO) terms by TGFβ and EGF. Further, only TGFβ induced the expression of epithelial to mesenchymal transition (EMT)-related proteins like matrix metalloproteinase 2 (MMP2). EGF, in contrast, made no major contribution to EMT marker expression on either the protein or the transcript level. In line with these expression patterns, TGFβ treatment significantly increased the invasive capacity of A549 cells, while EGF treatment did not. Moreover, the addition of EGF failed to enhance TGFβ-induced invasion. Overall, these data suggest that TGFβ and EGF can partly compensate for each other for stimulation of cell migration, but abrogation of TGFβ signaling may be more suitable to suppress cell invasion.
肿瘤及其微环境会产生多种生长因子,这些生长因子通过各种信号转导途径影响癌细胞的行为。生长因子,如转化生长因子β(TGFβ)和表皮生长因子(EGF),已被证明在不同的细胞模型中可诱导增殖、迁移和侵袭。这两种因子在癌症中经常过度表达,且常常共同发挥作用。尽管这两种因子在临床肿瘤学中都被视为合理的治疗靶点,但其对癌细胞迁移和侵袭的贡献的异同尚未完全明确。在此,我们通过应用视频显微镜、功能测定、免疫印迹、实时PCR和蛋白质组学,比较了用TGFβ、EGF以及两者联合处理A549肺腺癌细胞的影响。两种因子处理均能在相似程度上刺激A549细胞迁移,但动力学不同。联合处理具有相加效应。EGF诱导的迁移依赖于丝裂原活化蛋白激酶(MAPK)途径的激活。然而,尽管TGFβ能强烈激活该途径,但该途径对于TGFβ诱导的迁移并非必需。蛋白质组分析(数据可通过ProteomeXchange获取,标识符为PXD023024)显示,TGFβ和EGF诱导的迁移相关蛋白的表达模式以及相关的基因本体(GO)术语存在重叠。此外,只有TGFβ能诱导上皮-间质转化(EMT)相关蛋白如基质金属蛋白酶2(MMP2)的表达。相比之下,EGF在蛋白质或转录水平上对EMT标志物的表达没有显著贡献。与这些表达模式一致,TGFβ处理显著增加了A549细胞的侵袭能力,而EGF处理则没有。此外,添加EGF未能增强TGFβ诱导的侵袭。总体而言,这些数据表明,TGFβ和EGF在刺激细胞迁移方面可部分相互补偿,但消除TGFβ信号可能更适合抑制细胞侵袭。