RT info:eu-repo/semantics/article
T1 On-line surface plasmon resonance biosensing of vascular endothelial growth factor signaling in intact-human hepatoma cell lines
A1 Mauriz García, Elba
A1 Carbajo Pescador, Sara
A1 Ordoñez, Raquel
A1 García Fernández, María del Camino
A1 Mauriz Gutiérrez, José Luis
A1 Lechuga, Laura María
A1 González Gallego, Javier
A2 Enfermeria
K1 Enfermería
K1 Biodetección
K1 Resonancia de plasmón
K1 Endotelio
K1 Hepatoma
K1 3205 Medicina Interna
K1 3212 Salud Publica
K1 enferme
AB [EN] 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 x 105 cells mL1) 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 VEGFR inhibitor, CBOP11. 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 (24 x 104 cells mL1) onto a sensor surface previously immobilized with VEGF via thiol selfassembled 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 last 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.
PB Royal Society of Chemistry
SN 0003-2654
LK https://hdl.handle.net/10612/20041
UL https://hdl.handle.net/10612/20041
NO Mauriz, Carbajo-Pescador, Ordoñez, García-Fernández, Lechuga, Lechuga, & González-Gallego. (2014). On-line surface plasmon resonance biosensing of vascular endothelial growth factor signaling in intact-human hepatoma cell lines. Analyst, 139(6), 1426-1435. https://doi.org/10.1039/C3AN02211K
DS BULERIA. Repositorio Institucional de la Universidad de León
RD 07-jun-2024