Issue 11, 2019

Profiling protein–protein interactions of single cancer cells with in situ lysis and co-immunoprecipitation

Abstract

Heterogeneity in a tumor allows a small portion of cancer cells to survive and regrow upon targeted cancer therapy, eventually leading to cancer relapse. Such drug-resistant cells often exhibit dynamic adaptation of their signaling pathways at the level of protein–protein interactions (PPIs). To probe the rewiring of signaling pathways and the heterogeneity across individual cancer cells, we developed a single-cell version of the co-immunoprecipitation (co-IP) analysis that examines the amount and PPIs of target proteins immunoprecipitated from individual cells. The method captures cancer cells at predefined locations using a microfluidic chip, pulls down target proteins on the surface using antibodies, and lyses the captured cells in situ. Then, subsequent addition of eGFP-labeled downstream proteins enables the determination of the corresponding PPIs for the minimal amount of target proteins sampled from a single cell. We applied the technique to probe epidermal growth factor receptors (EGFRs) in PC9 lung adenocarcinoma cells. The results reveal that the strength of EGFR PPIs can be largely uncorrelated with the expression level of EGFRs in single cells. In addition, the individual PC9 cells showed markedly different patterns of PPIs, indicating a high heterogeneity in EGFR signaling within a genetically homogeneous population.

Graphical abstract: Profiling protein–protein interactions of single cancer cells with in situ lysis and co-immunoprecipitation

Supplementary files

Article information

Article type
Communication
Submitted
11 فرؤری 2019
Accepted
01 میٔ 2019
First published
01 میٔ 2019

Lab Chip, 2019,19, 1922-1928

Profiling protein–protein interactions of single cancer cells with in situ lysis and co-immunoprecipitation

J. Y. Ryu, J. Kim, M. J. Shon, J. Sun, X. Jiang, W. Lee and T. Yoon, Lab Chip, 2019, 19, 1922 DOI: 10.1039/C9LC00139E

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