Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/320126
Type: Artigo de Periódico
Title: 3d Cell-selex: Development Of Rna Aptamers As Molecular Probes For Pc-3 Tumor Cell Line
Author: Souza
AG; Marangoni
K; Fujimura
PT; Alves
PT; Silva
MJ; Bastos
VAF; Goulart
LR; Goulart
VA
Abstract: Human prostate cancer (PCa) is a highly heterogeneous and multifactorial disease. Current clinical biomarkers are not sufficiently accurate, thus being unable to predict the clinical outcome. Therefore, searching for new biomarkers aiming to improve diagnosis, prognosis and therapy is still required. In this study, we performed 3D Cell-SELEX against PC-3 prostate cancer cell line, a novel strategy to select specific nucleic acid ligands against spheroid cells in 3D cell culture. This original system combines Cell-SELEX, a process that exploits the cellular structure to generate specific ligands, and 3D cell culture, an approach that mimics the tissue microenvironment in vitro. In the first round of 3D Cell-SELEX, a negative selection against RWPE-1, non-tumor cell line, was performed to subtract non-tumor specific aptamers. The supernatant was used in eight additional rounds of selection, which were performed against PC-3 cell line. After nine selection cycles, eight PC-3 specific RNA aptamers were selected and sequenced. The aptamers presented sizes between 20 and 50 nucleotides-long, with low free energy (Delta G < - 13.6), which contributed for their spontaneous folding and high stability. Furthermore, our results showed the aptamer A4 as a specific ligand to prostate tumor cells, with dissociation constant in the nanomolar scale. Therefore, the novel 3D Cell-SELEX procedure improved the selection of PCa cell-surface ligands and the aptamer A4 has shown potential for the identification of prostate tumor cells, suggesting the application of this molecule in further screening assays for PCa. (C) 2016 Elsevier Inc. All rights reserved.
Subject: Prostate Cancer
3d Cell Culture
Cell-selex
Rna Aptamers
Editor: ELSEVIER INC
Rights: fechado
Identifier DOI: 10.1016/j.yexcr.2016.01.015
Address: http://www-sciencedirect-com.ez88.periodicos.capes.gov.br/science/article/pii/S0014482716300155
Date Issue: 2016
Appears in Collections:Unicamp - Artigos e Outros Documentos

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