Single-Cell Probe Force Studies to Identify Sox2 Overexpression-Promoted Cell Adhesion in MCF7 Breast Cancer Cells
The replacement of the cantilever tip by a living cell in Atomic Force Microscopy (AFM) experiments permits the direct quantification of cell–substrate and cell–cell adhesion forces. This single-cell probe force measurement technique, when complemented by microscopy, allows controlled manipulation o...
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| Format: | Article |
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MDPI AG
2020-04-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/9/4/935 |
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| author | Jagoba Iturri Andreas Weber María d.M. Vivanco José L. Toca-Herrera |
| author_facet | Jagoba Iturri Andreas Weber María d.M. Vivanco José L. Toca-Herrera |
| author_sort | Jagoba Iturri |
| collection | DOAJ |
| description | The replacement of the cantilever tip by a living cell in Atomic Force Microscopy (AFM) experiments permits the direct quantification of cell–substrate and cell–cell adhesion forces. This single-cell probe force measurement technique, when complemented by microscopy, allows controlled manipulation of the cell with defined location at the area of interest. In this work, a setup based on two glass half-slides, a non-fouling one with bacterial S-layer protein SbpA from <i>L. sphaericus</i> CMM 2177 and the second with a fibronectin layer, has been employed to measure the adhesion of MCF7 breast cancer cells to fibronectin films (using SbpA as control) and to other cells (symmetric vs. asymmetric systems). The measurements aimed to characterize and compare the adhesion capacities of parental cells and cells overexpressing the embryonic transcription factor Sox2, which have a higher capacity for invasion and are more resistant to endocrine therapy in vivo. Together with the use of fluorescence techniques (epifluorescence, Total Internal Fluorescence Microscopy (TIRF)), the visualization of vinculin and actin distribution in cells in contact with fibronectin surfaces is enabled, facilitating the monitoring and quantification of the formation of adhesion complexes. These findings demonstrate the strength of this combined approach to assess and compare the adhesion properties of cell lines and to illustrate the heterogeneity of adhesive strength found in breast cancer cells. |
| first_indexed | 2024-03-10T20:33:30Z |
| format | Article |
| id | doaj.art-ffe19b255ff84c868811a784c6317e59 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T20:33:30Z |
| publishDate | 2020-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-ffe19b255ff84c868811a784c6317e592023-11-19T21:16:51ZengMDPI AGCells2073-44092020-04-019493510.3390/cells9040935Single-Cell Probe Force Studies to Identify Sox2 Overexpression-Promoted Cell Adhesion in MCF7 Breast Cancer CellsJagoba Iturri0Andreas Weber1María d.M. Vivanco2José L. Toca-Herrera3Department of Nanobiotechnology (DNBT), Institute for Biophysics, BOKU University for Natural Resources and Life Sciences, Muthgasse 11 (Simon Zeisel Haus), A-1190 Vienna, AustriaDepartment of Nanobiotechnology (DNBT), Institute for Biophysics, BOKU University for Natural Resources and Life Sciences, Muthgasse 11 (Simon Zeisel Haus), A-1190 Vienna, AustriaCancer Heterogeneity Lab, Center for Cooperative Research in Biosciences (CIC bioGUNE), Basque Research and Technology Alliance (BRTA), Bizkaia Technology Park, 48160 Derio, SpainDepartment of Nanobiotechnology (DNBT), Institute for Biophysics, BOKU University for Natural Resources and Life Sciences, Muthgasse 11 (Simon Zeisel Haus), A-1190 Vienna, AustriaThe replacement of the cantilever tip by a living cell in Atomic Force Microscopy (AFM) experiments permits the direct quantification of cell–substrate and cell–cell adhesion forces. This single-cell probe force measurement technique, when complemented by microscopy, allows controlled manipulation of the cell with defined location at the area of interest. In this work, a setup based on two glass half-slides, a non-fouling one with bacterial S-layer protein SbpA from <i>L. sphaericus</i> CMM 2177 and the second with a fibronectin layer, has been employed to measure the adhesion of MCF7 breast cancer cells to fibronectin films (using SbpA as control) and to other cells (symmetric vs. asymmetric systems). The measurements aimed to characterize and compare the adhesion capacities of parental cells and cells overexpressing the embryonic transcription factor Sox2, which have a higher capacity for invasion and are more resistant to endocrine therapy in vivo. Together with the use of fluorescence techniques (epifluorescence, Total Internal Fluorescence Microscopy (TIRF)), the visualization of vinculin and actin distribution in cells in contact with fibronectin surfaces is enabled, facilitating the monitoring and quantification of the formation of adhesion complexes. These findings demonstrate the strength of this combined approach to assess and compare the adhesion properties of cell lines and to illustrate the heterogeneity of adhesive strength found in breast cancer cells.https://www.mdpi.com/2073-4409/9/4/935single-cell probeAFMepifluorescenceTIRFcell adhesionMCF7 cells |
| spellingShingle | Jagoba Iturri Andreas Weber María d.M. Vivanco José L. Toca-Herrera Single-Cell Probe Force Studies to Identify Sox2 Overexpression-Promoted Cell Adhesion in MCF7 Breast Cancer Cells Cells single-cell probe AFM epifluorescence TIRF cell adhesion MCF7 cells |
| title | Single-Cell Probe Force Studies to Identify Sox2 Overexpression-Promoted Cell Adhesion in MCF7 Breast Cancer Cells |
| title_full | Single-Cell Probe Force Studies to Identify Sox2 Overexpression-Promoted Cell Adhesion in MCF7 Breast Cancer Cells |
| title_fullStr | Single-Cell Probe Force Studies to Identify Sox2 Overexpression-Promoted Cell Adhesion in MCF7 Breast Cancer Cells |
| title_full_unstemmed | Single-Cell Probe Force Studies to Identify Sox2 Overexpression-Promoted Cell Adhesion in MCF7 Breast Cancer Cells |
| title_short | Single-Cell Probe Force Studies to Identify Sox2 Overexpression-Promoted Cell Adhesion in MCF7 Breast Cancer Cells |
| title_sort | single cell probe force studies to identify sox2 overexpression promoted cell adhesion in mcf7 breast cancer cells |
| topic | single-cell probe AFM epifluorescence TIRF cell adhesion MCF7 cells |
| url | https://www.mdpi.com/2073-4409/9/4/935 |
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