Characterization of MNK1b DNA Aptamers That Inhibit Proliferation in MDA-MB231 Breast Cancer Cells
| Autor: | M. Elena Martín, Marta García-Hernández, Gerónimo Fernández, Eva M. García-Recio, Celia Pinto-Díez, Víctor M. González, M. Isabel Pérez-Morgado |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Aptamer Population MNK1 translation Biology 03 medical and health sciences Drug Discovery cancer education education.field_of_study Kinase Oligonucleotide SELEX EIF4E lcsh:RM1-950 aptamer Molecular biology 030104 developmental biology Real-time polymerase chain reaction lcsh:Therapeutics. Pharmacology Mitogen-activated protein kinase biology.protein Molecular Medicine Original Article Systematic evolution of ligands by exponential enrichment |
| Zdroj: | Molecular Therapy: Nucleic Acids, Vol 5, Iss C (2016) Repositorio Institucional de la Consejería de Sanidad de la Comunidad de Madrid Consejería de Sanidad de la Comunidad de Madrid Molecular Therapy. Nucleic Acids |
| ISSN: | 2162-2531 |
| Popis: | Elevated expression levels of eukaryotic initiation factor 4E (eIF4E) promote cancer development and progression. MAP kinase interacting kinases (MNKs) modulate the function of eIF4E through the phosphorylation that is necessary for oncogenic transformation. Therefore, pharmacologic MNK inhibitors may provide a nontoxic and effective anticancer strategy. MNK1b is a truncated isoform of MNK1a that is active in the absence of stimuli. Using in vitro selection, high-affinity DNA aptamers to MNK1b were selected from a library of ssDNA. Selection was monitored using the enzyme-linked oligonucleotide assay (ELONA), and the selected aptamer population was cloned and sequenced. Four groups of aptamers were identified, and the affinities of one representative for rMNK1b were determined using ELONA and quantitative polymerase chain reaction. Two aptamers, named apMNK2F and apMNK3R, had a lower Kd in the nmol/l range. The secondary structure of the selected aptamers was predicted using mFold, and the QGRS Mapper indicated the presence of potential G-quadruplex structures in both aptamers. The selected aptamers were highly specific against MNK1, showing higher affinity to MNK1b than to MNK1a. Interestingly, both aptamers were able to produce significant translation inhibition and prevent tumor cell proliferation and migration and colony formation in breast cancer cells. These results indicate that MNK1 aptamers have an attractive therapeutic potential. |
| Databáze: | OpenAIRE |
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