Department of Chemistry, Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Tsinghua University, Beijing 100084, China.
Department of Electrical and Systems Engineering, Washington University in St Louis, MO 63130, USA.
Analyst. 2018 Nov 7;143(21):5264-5270. doi: 10.1039/c8an01534a. Epub 2018 Oct 3.
Epidermal growth factor receptor (EGFR), which belongs to the second-largest protein family for cell signal transduction, plays crucial roles in homeostasis, cellular organized patterns and most human cancers. In EGFR-activated signaling networks, the detection of the spatial and temporal dynamics of cascades that encode the many cell fates is still a challenge. Here, we report real-time imaging of epidermal growth factor (EGF)-induced EGFR activation and its signaling cascade in single A431 cells using surface plasmon resonance (SPR) microscopy. A two-phase SPR response pattern was observed within 30 min after EGF treatment, including a positive SPR response that was related to the EGFR-activated mass redistribution in the first 600 s, and a subsequent negative SPR signal caused by the morphological change of the cells. Furthermore, the inhibitor analysis verified that AG1478 inhibited the response from the whole the cell, whereas cytochalasin B strongly inhibited the response from the cell edge region.
表皮生长因子受体(EGFR)属于细胞信号转导的第二大蛋白家族,在维持内环境稳定、细胞组织形态和大多数人类癌症中发挥着关键作用。在 EGFR 激活的信号网络中,检测编码多种细胞命运的级联反应的时空动态仍然是一个挑战。在这里,我们使用表面等离子体共振(SPR)显微镜实时成像表皮生长因子(EGF)诱导的 EGFR 激活及其信号级联在单个 A431 细胞中的作用。在 EGF 处理后 30 分钟内观察到两阶段 SPR 响应模式,包括与前 600 秒 EGFR 激活导致的质量重分布相关的正 SPR 响应,以及随后由细胞形态变化引起的负 SPR 信号。此外,抑制剂分析验证了 AG1478 抑制了整个细胞的反应,而细胞松弛素 B 强烈抑制了细胞边缘区域的反应。