On-line surface plasmon resonance biosensing of vascular endothelial growth factor signaling in intact-human hepatoma cell lines
Mauriz, Elba (Universidad de León)
Carbajo Pescador, Sara (Universidad de León)
Ordóñez, Raquel (Universidad de León)
García Fernández, M. C. (Universidad de León)
Mauriz, José L. (Universidad de León)
Lechuga, Laura M (Institut Català de Nanociència i Nanotecnologia)
González-Gallego, Javier (Universidad de León)

Date:	2014
Abstract:	Surface plasmon resonance (SPR) monitoring of biorecognition events at intracellular levels is a valuable tool for studying the angiogenic response of carcinoma living cells during tumor growth and proliferation. We report here a comparative study of two different strategies to detect human hepatoma cell interactions between transmembrane vascular endothelial growth factor receptor (VEGFR2) and vascular endothelial growth factor (VEGF). To monitor VEGFR2 activation after VEGF stimulation, intact hepatocellular carcinoma HepG2 or Huh7 cells (2 × 10 cells per mL) were directly immobilized on the sensor chip. Distinguishable SPR sensorgrams were obtained for each cell line depending on the time required for VEGFR2 activation. SPR signals for VEGF-VEGFR2 binding were inhibited by the VEGFR inhibitor, CBO-P11. The SPR response after VEGF stimulation/inhibition was in good agreement with the results observed by immunoblotting analysis. In a second approach we used intact cell lines as analytes. SPR analysis was done by injecting HepG2 and HuH7 cell suspensions (2-4 × 10 cells per mL) onto a sensor surface previously immobilized with VEGF via a thiol self-assembled monolayer (SAM). Specificity and reproducibility were evaluated reusing the same chip surface over more than 60 complete regeneration cycles. Comparison between both methods yielded differences in terms of reliability, making the latter strategy more effective for the analysis of real samples. The investigation of VEGF signaling in intact human hepatoma living cells by SPR monitoring comprises a novel and promising design for the study of tumor angiogenesis via downregulation of VEGF and VEGFR2 pathways. Further investigation on VEGFR activation and vascular function could contribute to establish a robust and meaningful tool for early cancer diagnostics.
Rights:	Tots els drets reservats.
Language:	Anglès
Document:	Article ; recerca ; Versió acceptada per publicar
Subject:	Carcinoma, Hepatocellular ; Cell Line, Tumor ; Equipment Design ; Hep G2 Cells ; Humans ; Liver Neoplasms ; Reproducibility of Results ; Signal Transduction ; Surface Plasmon Resonance ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor Receptor-2
Published in:	Analyst, Vol. 139, Issue 6 (March 2014) , p. 1426-1435, ISSN 1364-5528

DOI: 10.1039/c3an02211k

Postprint 37 p, 1.4 MB

The record appears in these collections:
Research literature > UAB research groups literature > Research Centres and Groups (research output) > Experimental sciences > Catalan Institute of Nanoscience and Nanotechnology (ICN2)
Articles > Research articles
Articles > Published articles