Comprehensive analysis of signal transduction in three-dimensional ECM-based tumor cell cultures

¹ OncoRay – National Center for Radiation Research in Oncology, Medical Faculty Carl Gustav Carus, Dresden University of Technology, 01307 Dresden, Germany

² Radiation Oncology Branch, Center for Cancer Research,National Institutes of Health/National Cancer Institute, Bethesda, MD 20892, USA

³ Department of Radiotherapy and Oncology, University of Frankfurt, 60590 Frankfurt am Main, Germany

⁴ Full Moon BioSystems Inc., Sunnyvale, CA 94085, USA

⁵ Department of Radiation Oncology, University Hospital and Medical Faculty Carl Gustav Carus, Dresden University of Technology, 01307 Dresden, Germany

⁶ Helmholtz Center Dresden-Rossendorf, Institute of Radiooncology, 01328 Dresden, Germany

⁷ German Cancer Consortium (DKTK), 01307 Dresden, Germany

⁸ German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany

JBM 2015 , 2(4), 1;

Published: 17 November 2015

© 2015 by the author. Licensee POL Scientific, USA. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )

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Abstract

Analysis of signal transduction and protein phosphorylation is fundamental to understand physiological and pathological cell behavior as well as identification of novel therapeutic targets. Despite the fact that more physiological three-dimensional cell culture assays are increasingly used, particularly proteomics and phosphoproteomics remain challenging due to easy, robust and reproducible sample preparation. Here, we present an easy-to-perform, reliable and time-efficient method for the production of 3D cell lysates without compromising cell adhesion before cell lysis. The samples can be used for Western blotting as well as phosphoproteome array technology. This technique would be of interest for researchers working in all fields of biology and drug development.

Keywords

3D cell culture

extracellular matrix

protein analysis

phosphoproteomics

signaling

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