Tight association of autophagy and cell cycle in leukemia cells
Abstract Background Autophagy plays an essential role in maintaining cellular homeostasis and in the response to cellular stress. Autophagy is also involved in cell cycle progression, yet the relationship between these processes is not clearly defined. Results In exploring this relationship, we obse...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-04-01
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| Series: | Cellular & Molecular Biology Letters |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s11658-022-00334-8 |
| _version_ | 1811291024370171904 |
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| author | Alena Gschwind Christian Marx Marie D. Just Paula Severin Hannah Behring Lisa Marx-Blümel Sabine Becker Linda Rothenburger Martin Förster James F. Beck Jürgen Sonnemann |
| author_facet | Alena Gschwind Christian Marx Marie D. Just Paula Severin Hannah Behring Lisa Marx-Blümel Sabine Becker Linda Rothenburger Martin Förster James F. Beck Jürgen Sonnemann |
| author_sort | Alena Gschwind |
| collection | DOAJ |
| description | Abstract Background Autophagy plays an essential role in maintaining cellular homeostasis and in the response to cellular stress. Autophagy is also involved in cell cycle progression, yet the relationship between these processes is not clearly defined. Results In exploring this relationship, we observed that the inhibition of autophagy impaired the G2/M phase-arresting activity of etoposide but enhanced the G1 phase-arresting activity of palbociclib. We further investigated the connection of basal autophagy and cell cycle by utilizing the autophagosome tracer dye Cyto-ID in two ways. First, we established a double-labeling flow-cytometric procedure with Cyto-ID and the DNA probe DRAQ5, permitting the cell cycle phase-specific determination of autophagy in live cells. This approach demonstrated that different cell cycle phases were associated with different autophagy levels: G1-phase cells had the lowest level, and G2/M-phase cells had the highest one. Second, we developed a flow-cytometric cell-sorting procedure based on Cyto-ID that separates cell populations into fractions with low, medium, and high autophagy. Cell cycle analysis of Cyto-ID-sorted cells confirmed that the high-autophagy fraction contained a much higher percentage of G2/M-phase cells than the low-autophagy fraction. In addition, Cyto-ID-based cell sorting also proved to be useful for assessing other autophagy-related processes: extracellular flux analysis revealed metabolic differences between the cell populations, with higher autophagy being associated with higher respiration, higher mitochondrial ATP production, and higher glycolysis. Conclusion This work provides clear evidence of high autophagy in G2/M-phase cells by establishing a novel cell sorting technique based on Cyto-ID. |
| first_indexed | 2024-04-13T04:23:10Z |
| format | Article |
| id | doaj.art-80c924d65a344261b247c7fdff57d0f2 |
| institution | Directory Open Access Journal |
| issn | 1425-8153 1689-1392 |
| language | English |
| last_indexed | 2024-04-13T04:23:10Z |
| publishDate | 2022-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Cellular & Molecular Biology Letters |
| spelling | doaj.art-80c924d65a344261b247c7fdff57d0f22022-12-22T03:02:39ZengBMCCellular & Molecular Biology Letters1425-81531689-13922022-04-0127112010.1186/s11658-022-00334-8Tight association of autophagy and cell cycle in leukemia cellsAlena Gschwind0Christian Marx1Marie D. Just2Paula Severin3Hannah Behring4Lisa Marx-Blümel5Sabine Becker6Linda Rothenburger7Martin Förster8James F. Beck9Jürgen Sonnemann10Department of Pediatric Hematology and Oncology, Children’s Clinic, Jena University HospitalLeibniz Institute on Aging-Fritz Lipmann Institute (FLI)Department of Pediatric Hematology and Oncology, Children’s Clinic, Jena University HospitalDepartment of Pediatric Hematology and Oncology, Children’s Clinic, Jena University HospitalDepartment of Pediatric Hematology and Oncology, Children’s Clinic, Jena University HospitalDepartment of Pediatric Hematology and Oncology, Children’s Clinic, Jena University HospitalDepartment of Pediatric Hematology and Oncology, Children’s Clinic, Jena University HospitalLeibniz Institute on Aging-Fritz Lipmann Institute (FLI)Clinic of Internal Medicine I, Jena University HospitalDepartment of Pediatric Hematology and Oncology, Children’s Clinic, Jena University HospitalDepartment of Pediatric Hematology and Oncology, Children’s Clinic, Jena University HospitalAbstract Background Autophagy plays an essential role in maintaining cellular homeostasis and in the response to cellular stress. Autophagy is also involved in cell cycle progression, yet the relationship between these processes is not clearly defined. Results In exploring this relationship, we observed that the inhibition of autophagy impaired the G2/M phase-arresting activity of etoposide but enhanced the G1 phase-arresting activity of palbociclib. We further investigated the connection of basal autophagy and cell cycle by utilizing the autophagosome tracer dye Cyto-ID in two ways. First, we established a double-labeling flow-cytometric procedure with Cyto-ID and the DNA probe DRAQ5, permitting the cell cycle phase-specific determination of autophagy in live cells. This approach demonstrated that different cell cycle phases were associated with different autophagy levels: G1-phase cells had the lowest level, and G2/M-phase cells had the highest one. Second, we developed a flow-cytometric cell-sorting procedure based on Cyto-ID that separates cell populations into fractions with low, medium, and high autophagy. Cell cycle analysis of Cyto-ID-sorted cells confirmed that the high-autophagy fraction contained a much higher percentage of G2/M-phase cells than the low-autophagy fraction. In addition, Cyto-ID-based cell sorting also proved to be useful for assessing other autophagy-related processes: extracellular flux analysis revealed metabolic differences between the cell populations, with higher autophagy being associated with higher respiration, higher mitochondrial ATP production, and higher glycolysis. Conclusion This work provides clear evidence of high autophagy in G2/M-phase cells by establishing a novel cell sorting technique based on Cyto-ID.https://doi.org/10.1186/s11658-022-00334-8AutophagyCell cycleCell sortingCyto-IDDRAQ5Metabolic analysis |
| spellingShingle | Alena Gschwind Christian Marx Marie D. Just Paula Severin Hannah Behring Lisa Marx-Blümel Sabine Becker Linda Rothenburger Martin Förster James F. Beck Jürgen Sonnemann Tight association of autophagy and cell cycle in leukemia cells Cellular & Molecular Biology Letters Autophagy Cell cycle Cell sorting Cyto-ID DRAQ5 Metabolic analysis |
| title | Tight association of autophagy and cell cycle in leukemia cells |
| title_full | Tight association of autophagy and cell cycle in leukemia cells |
| title_fullStr | Tight association of autophagy and cell cycle in leukemia cells |
| title_full_unstemmed | Tight association of autophagy and cell cycle in leukemia cells |
| title_short | Tight association of autophagy and cell cycle in leukemia cells |
| title_sort | tight association of autophagy and cell cycle in leukemia cells |
| topic | Autophagy Cell cycle Cell sorting Cyto-ID DRAQ5 Metabolic analysis |
| url | https://doi.org/10.1186/s11658-022-00334-8 |
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