Single-cell proteomic chip for profiling intracellular signaling pathways in single tumor cells
Single-cell proteomic chip for profiling intracellular signaling pathways in single tumor cells
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DOI：
10.1073/pnas.1110865109
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发表时间：
2012-01-10
影响因子：
11.1
通讯作者：
Heath, James R.
中科院分区：
文献类型：
--
作者：
Shi, Qihui;Qin, Lidong;Heath, James R.
我们描述了一种微芯片，旨在量化单细胞的十几种细胞质和膜蛋白的水平。我们使用该平台来评估与EGF受体介导的PI 3 K信号通路相关的蛋白质-蛋白质相互作用。通过在2 nL体积的室中分离限定数量的细胞（n = 0-5）来实现单细胞灵敏度，每个室用微型抗体阵列的两个拷贝进行图案化。将细胞在芯片上裂解，并使用抗体阵列测定释放的蛋白质的水平。我们研究了三个代表癌症多形性胶质母细胞瘤的同基因细胞系，在基础水平，在EGF刺激下，以及在厄洛替尼抑制加EGF刺激下。测量的蛋白质丰度与以前的工作是一致的，单细胞分析独特地揭示了单细胞异质性，以及不同类型和强度的蛋白质-蛋白质相互作用。该平台有助于全面了解肿瘤细胞中改变的信号转导网络，并深入了解靶向治疗对蛋白质信号转导网络的影响。
We describe a microchip designed to quantify the levels of a dozen cytoplasmic and membrane proteins from single cells. We use the platform to assess protein-protein interactions associated with the EGF-receptor-mediated PI3K signaling pathway. Single-cell sensitivity is achieved by isolating a defined number of cells (n = 0-5) in 2 nL volume chambers, each of which is patterned with two copies of a miniature antibody array. The cells are lysed on-chip, and the levels of released proteins are assayed using the antibody arrays. We investigate three isogenic cell lines representing the cancer glioblastoma multiforme, at the basal level, under EGF stimulation, and under erlotinib inhibition plus EGF stimulation. The measured protein abundances are consistent with previous work, and single-cell analysis uniquely reveals single-cell heterogeneity, and different types and strengths of protein-protein interactions. This platform helps provide a comprehensive picture of altered signal transduction networks in tumor cells and provides insight into the effect of targeted therapies on protein signaling networks.