These authors contributed equally to this work.
Application of two-dimensional gel-based mass spectrometry to functionally dissect resistance to targeted cancer therapy
Article first published online: 10 DEC 2013
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
PROTEOMICS - Clinical Applications
Special Issue: Focus on Diagnostic Proteomics
Volume 7, Issue 11-12, pages 813–824, December 2013
How to Cite
Klein, O., Rohwer, N., de Molina, K. F., Mergler, S., Wessendorf, P., Herrmann, M., Klose, J. and Cramer, T. (2013), Application of two-dimensional gel-based mass spectrometry to functionally dissect resistance to targeted cancer therapy. Prot. Clin. Appl., 7: 813–824. doi: 10.1002/prca.201300056
Colour Online: See the article online to view Figs. 1, 4 and 6 in colour.
- Issue published online: 10 DEC 2013
- Article first published online: 10 DEC 2013
- Accepted manuscript online: 31 OCT 2013 02:58AM EST
- Manuscript Accepted: 5 OCT 2013
- Manuscript Revised: 1 OCT 2013
- Manuscript Received: 3 JUL 2013
- Deutsche Forschungsgemeinschaft (DFG)
- Deutsche Krebshilfe
- Bundesministerium für Bildung und Forschung (BMBF)
- German Consortium for Translational Cancer Research (DKTK)
- Berlin-Brandenburg Center for Regenerative Therapies
- Gastric cancer;
- Targeted therapy;
- Therapy resistance
The majority of gastric cancers are diagnosed at advanced stages, characterized by robust therapy resistance. The oncoprotein hypoxia-inducible factor 1 (HIF-1) is associated with therapy resistance, partly via activation of the DNA damage response. We have noted a robust ability of gastric cancer cells to functionally compensate the loss of HIF-1 in vitro. The purpose of this study was to identify molecular pathways that underlie this compensation.
We performed 2DE to compare the nuclear proteome of wild-type and HIF-1-deficient gastric cancer cells. Differently expressed protein spots were identified via MS). After bioinformatic evaluation, functional validation of selected identified pathways was performed.
2DE displayed a total of 2523 protein spots, from which 87 were identified as regulated by HIF-1. Seventy of the identified spots were different proteins and 17 were isoforms. Bioinformatic analyses revealed that a significant amount of the identified proteins were related to cellular survival pathways. Specifically, members of the proteasome pathway were found upregulated upon loss of HIF-1. Combined inhibition of HIF-1 and the proteasome inflicted significant DNA damage, supporting the hypothesis that the proteasome is of functional importance to compensate the loss of HIF-1.
Conclusions and clinical relevance
Our data show robust and functional changes of the nuclear proteome upon inactivation of the HIF-1 oncoprotein in gastric cancer cells. We propose that 2DE-MS represents a useful tool to functionally dissect resistance mechanisms to targeted therapy and to identify novel targets for antiproliferative combination therapy.