In situ monitoring of PTHLH secretion in neuroblastoma cells cultured onto nanoporous membranes
| Autor: | Carlos J. Rodríguez-Hernández, Carmen Torres, Marisol Espinoza-Castañeda, Alejandro Chamorro-Garcia, Arben Merkoçi, Alfredo de la Escosura-Muñiz |
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| Přispěvatelé: | Generalitat de Catalunya, Ministerio de Economía y Competitividad (España), European Commission, Ministerio de Sanidad, Consumo y Bienestar Social (España), Fundación Antonio Cabré |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Cancer cells Biomedical Engineering Biophysics Biosensing Techniques 02 engineering and technology Cell Line Nanopores/nanochannels Nanopores Neuroblastoma 03 medical and health sciences Settore CHIM/01 Cell Line Tumor Electrochemistry Humans Electrochemical detection Secretion Gene knockdown Parathyroid hormone-related protein Cell growth Chemistry Parathyroid Hormone-Related Protein Membranes Artificial Equipment Design General Medicine Transfection PTHLH 021001 nanoscience & nanotechnology 3. Good health Cell biology HaCaT 030104 developmental biology Cell culture Cancer cell 0210 nano-technology Biotechnology |
| Zdroj: | BIOSENSORS & BIOELECTRONICS r-FSJD. Repositorio Institucional de Producción Científica de la Fundació Sant Joan de Déu instname r-FSJD: Repositorio Institucional de Producción Científica de la Fundació Sant Joan de Déu Fundació Sant Joan de Déu Digital.CSIC. Repositorio Institucional del CSIC Dipòsit Digital de Documents de la UAB Universitat Autònoma de Barcelona |
| ISSN: | 0956-5663 |
| DOI: | 10.1016/j.bios.2018.01.064 |
| Popis: | In this work, we propose for the first time the use of anodic aluminum oxide (AAO) nanoporous membranes for in situ monitoring of parathyroid hormone-like hormone (PTHLH) secretion in cultured human cells. The biosensing system is based on the nanochannels blockage upon immunocomplex formation, which is electrically monitored through the voltammetric oxidation of Prussian blue nanoparticles (PBNPs). Models evaluated include a neuroblastoma cell line (SK-N-AS) and immortalized keratinocytes (HaCaT) as a control of high PTHLH production. The effect of total number of seeded cells and incubation time on the secreted PTHLH levels is assessed, finding that secreted PTHLH levels range from approximately 60 to 400 ng/mL. Moreover, our methodology is also applied to analyse PTHLH production following PTHLH gene knockdown upon transient cell transfection with a specific silencing RNA (siRNA). Given that inhibition of PTHLH secretion reduces cell proliferation, survival and invasiveness in a number of tumors, our system provides a powerful tool for the preclinical evaluation of therapies that regulate PTHLH production. This nanoporous membrane – based sensing technology might be useful to monitor the active secretion of other proteins as well, thus contributing to characterize their regulation and function. We acknowledge support from the Spanish MINECO under projects MAT2014-52485-P and PCIN-2016-066 (program Euronanomed 2) and from Secretaria d’Universitats i Recerca del Departament d’Economia i Coneixement de la Generalitat de Catalunya (Grant 2014 SGR 260). This work is also funded by the CERCA Programme/Generalitat de Catalunya. ICN2 is supported by the Severo Ochoa program from Spanish MINECO (Grant no. SEV-2013-0295). CdT's group is supported by the Spanish Ministry of Health (PI14/00040), Euronanomed 2 (AC16/00026), H2020-MSCA-ITN-2015 Marie Curie International Training Network (675228) and Fundació Antonio Cabré. |
| Databáze: | OpenAIRE |
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