Selective Targeting to Glioma with Nucleic Acid Aptamers
Malignant glioma is characterised by a rapid growth rate and high capacity for invasive infiltration to surrounding brain tissue; hence, diagnosis and treatment is difficult and patient survival is poor. Aptamers contribute a promising and unique technology for the in vitro imaging of live cells and...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Public Library of Science
2015
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4529171/ https://www.ncbi.nlm.nih.gov/pubmed/26252900 http://dx.doi.org/10.1371/journal.pone.0134957 |
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author | Aptekar, Shraddha Arora, Mohit Lawrence, Clare Louise Lea, Robert William Ashton, Katherine Dawson, Tim Alder, Jane Elizabeth Shaw, Lisa |
author_facet | Aptekar, Shraddha Arora, Mohit Lawrence, Clare Louise Lea, Robert William Ashton, Katherine Dawson, Tim Alder, Jane Elizabeth Shaw, Lisa |
author_sort | Aptekar, Shraddha |
collection | PubMed |
description | Malignant glioma is characterised by a rapid growth rate and high capacity for invasive infiltration to surrounding brain tissue; hence, diagnosis and treatment is difficult and patient survival is poor. Aptamers contribute a promising and unique technology for the in vitro imaging of live cells and tissues, with a potentially bright future in clinical diagnostics and therapeutics for malignant glioma. The binding selectivity, uptake capacity and binding target of two DNA aptamers, SA43 and SA44, were investigated in glioma cells and patient tissues. The binding assay showed that SA43 and SA44 bound with strong affinity (K(d), 21.56 ± 4.60 nM and K(d), 21.11 ± 3.30 nM respectively) to the target U87MG cells. Quantitative analysis by flow cytometry showed that the aptamers were able to actively internalise in U87MG and 1321N1 glioma cells compared to the non-cancerous and non-glioma cell types. Confocal microscopy confirmed staining in the cytoplasm, and co-localisation studies with endoplasmic reticulum, Golgi apparatus and lysosomal markers suggested internalisation and compartmentalisation within the endomembrane system. Both aptamers selectively bound to Ku 70 and Ku 80 DNA repair proteins as determined by aptoprecipitation (AP) followed by mass spectrometry analysis and confirmation by Western blot. In addition, aptohistochemical (AHC) staining on paraffin embedded, formalin fixed patient tissues revealed that the binding selectivity was significantly higher for SA43 aptamer in glioma tissues (grade I, II, III and IV) compared to the non-cancerous tissues, whereas SA44 did not show selectivity towards glioma tissues. The results indicate that SA43 aptamer can differentiate between glioma and non-cancerous cells and tissues and therefore, shows promise for histological diagnosis of glioma. |
format | Online Article Text |
id | pubmed-4529171 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-45291712015-08-12 Selective Targeting to Glioma with Nucleic Acid Aptamers Aptekar, Shraddha Arora, Mohit Lawrence, Clare Louise Lea, Robert William Ashton, Katherine Dawson, Tim Alder, Jane Elizabeth Shaw, Lisa PLoS One Research Article Malignant glioma is characterised by a rapid growth rate and high capacity for invasive infiltration to surrounding brain tissue; hence, diagnosis and treatment is difficult and patient survival is poor. Aptamers contribute a promising and unique technology for the in vitro imaging of live cells and tissues, with a potentially bright future in clinical diagnostics and therapeutics for malignant glioma. The binding selectivity, uptake capacity and binding target of two DNA aptamers, SA43 and SA44, were investigated in glioma cells and patient tissues. The binding assay showed that SA43 and SA44 bound with strong affinity (K(d), 21.56 ± 4.60 nM and K(d), 21.11 ± 3.30 nM respectively) to the target U87MG cells. Quantitative analysis by flow cytometry showed that the aptamers were able to actively internalise in U87MG and 1321N1 glioma cells compared to the non-cancerous and non-glioma cell types. Confocal microscopy confirmed staining in the cytoplasm, and co-localisation studies with endoplasmic reticulum, Golgi apparatus and lysosomal markers suggested internalisation and compartmentalisation within the endomembrane system. Both aptamers selectively bound to Ku 70 and Ku 80 DNA repair proteins as determined by aptoprecipitation (AP) followed by mass spectrometry analysis and confirmation by Western blot. In addition, aptohistochemical (AHC) staining on paraffin embedded, formalin fixed patient tissues revealed that the binding selectivity was significantly higher for SA43 aptamer in glioma tissues (grade I, II, III and IV) compared to the non-cancerous tissues, whereas SA44 did not show selectivity towards glioma tissues. The results indicate that SA43 aptamer can differentiate between glioma and non-cancerous cells and tissues and therefore, shows promise for histological diagnosis of glioma. Public Library of Science 2015-08-07 /pmc/articles/PMC4529171/ /pubmed/26252900 http://dx.doi.org/10.1371/journal.pone.0134957 Text en © 2015 Aptekar et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Aptekar, Shraddha Arora, Mohit Lawrence, Clare Louise Lea, Robert William Ashton, Katherine Dawson, Tim Alder, Jane Elizabeth Shaw, Lisa Selective Targeting to Glioma with Nucleic Acid Aptamers |
title | Selective Targeting to Glioma with Nucleic Acid Aptamers |
title_full | Selective Targeting to Glioma with Nucleic Acid Aptamers |
title_fullStr | Selective Targeting to Glioma with Nucleic Acid Aptamers |
title_full_unstemmed | Selective Targeting to Glioma with Nucleic Acid Aptamers |
title_short | Selective Targeting to Glioma with Nucleic Acid Aptamers |
title_sort | selective targeting to glioma with nucleic acid aptamers |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4529171/ https://www.ncbi.nlm.nih.gov/pubmed/26252900 http://dx.doi.org/10.1371/journal.pone.0134957 |
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