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Impact of Doxorubicin on Cell-Substrate Topology
Variable-Angle Total Internal Reflection Fluorescence Microscopy (VA-TIRFM) is applied in view of early detection of cellular responses to the cytostatic drug doxorubicin. Therefore, we determined cell-substrate topology of cultivated CHO cells transfected with a membrane-associated Green Fluorescen...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9181088/ https://www.ncbi.nlm.nih.gov/pubmed/35682954 http://dx.doi.org/10.3390/ijms23116277 |
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author | Krecsir, Andreas Richter, Verena Wagner, Michael Schneckenburger, Herbert |
author_facet | Krecsir, Andreas Richter, Verena Wagner, Michael Schneckenburger, Herbert |
author_sort | Krecsir, Andreas |
collection | PubMed |
description | Variable-Angle Total Internal Reflection Fluorescence Microscopy (VA-TIRFM) is applied in view of early detection of cellular responses to the cytostatic drug doxorubicin. Therefore, we determined cell-substrate topology of cultivated CHO cells transfected with a membrane-associated Green Fluorescent Protein (GFP) in the nanometer range prior to and subsequent to the application of doxorubicin. Cell-substrate distances increased up to a factor of 2 after 24 h of application. A reduction of these distances by again a factor 2 was observed upon cell aging, and an influence of the cultivation time is presently discussed. Applicability of VA-TIRFM was supported by measurements of MCF-7 breast cancer cells after membrane staining and incubation with doxorubicin, when cell-substrate distances increased again by a factor ≥ 2. So far, our method needs well-defined cell ages and staining of cell membranes or transfection with GFP or related molecules. Use of intrinsic fluorescence or even light-scattering methods to various cancer cell lines could make this method more universal in the future, e.g., in the context of early detection of apoptosis. |
format | Online Article Text |
id | pubmed-9181088 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-91810882022-06-10 Impact of Doxorubicin on Cell-Substrate Topology Krecsir, Andreas Richter, Verena Wagner, Michael Schneckenburger, Herbert Int J Mol Sci Communication Variable-Angle Total Internal Reflection Fluorescence Microscopy (VA-TIRFM) is applied in view of early detection of cellular responses to the cytostatic drug doxorubicin. Therefore, we determined cell-substrate topology of cultivated CHO cells transfected with a membrane-associated Green Fluorescent Protein (GFP) in the nanometer range prior to and subsequent to the application of doxorubicin. Cell-substrate distances increased up to a factor of 2 after 24 h of application. A reduction of these distances by again a factor 2 was observed upon cell aging, and an influence of the cultivation time is presently discussed. Applicability of VA-TIRFM was supported by measurements of MCF-7 breast cancer cells after membrane staining and incubation with doxorubicin, when cell-substrate distances increased again by a factor ≥ 2. So far, our method needs well-defined cell ages and staining of cell membranes or transfection with GFP or related molecules. Use of intrinsic fluorescence or even light-scattering methods to various cancer cell lines could make this method more universal in the future, e.g., in the context of early detection of apoptosis. MDPI 2022-06-03 /pmc/articles/PMC9181088/ /pubmed/35682954 http://dx.doi.org/10.3390/ijms23116277 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Communication Krecsir, Andreas Richter, Verena Wagner, Michael Schneckenburger, Herbert Impact of Doxorubicin on Cell-Substrate Topology |
title | Impact of Doxorubicin on Cell-Substrate Topology |
title_full | Impact of Doxorubicin on Cell-Substrate Topology |
title_fullStr | Impact of Doxorubicin on Cell-Substrate Topology |
title_full_unstemmed | Impact of Doxorubicin on Cell-Substrate Topology |
title_short | Impact of Doxorubicin on Cell-Substrate Topology |
title_sort | impact of doxorubicin on cell-substrate topology |
topic | Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9181088/ https://www.ncbi.nlm.nih.gov/pubmed/35682954 http://dx.doi.org/10.3390/ijms23116277 |
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