Radiosensitivity of Early and Late M-Phase HeLa Cells Isolated by a Combination of Fluorescent Ubiquitination-Based Cell Cycle Indicator (Fucci) and Mitotic Shake-Off
| Autor: | Kengo Sakaguchi, Masahiko Miura, Shifumi Deguchi, Motoshi Nakayama, Atsushi Kaida |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Genetics
Radiation Cell division medicine.diagnostic_test Ubiquitination Biophysics Mitosis Cell cycle Biology biology.organism_classification Radiation Tolerance Fluorescence Cell biology Flow cytometry HeLa Radioresistance medicine Humans Radiology Nuclear Medicine and imaging Radiosensitivity Cell Division HeLa Cells |
| Zdroj: | Radiation Research. 176:407-411 |
| ISSN: | 1938-5404 0033-7587 |
| DOI: | 10.1667/rr2608.1 |
| Popis: | The mitotic shake-off method revealed the remarkable variation of radiosensitivity of HeLa cells during the cell cycle: M phase shows the greatest radiosensitivity and late S phase the greatest radioresistance. This method harvests all M-phase cells with a round shape, making it impossible to further subdivide M-phase cells. Recently, the fluorescent ubiquitination-based cell cycle indicator (Fucci) was developed; this system basically causes cells in G(1) to emit red fluorescence and other cells to emit green fluorescence. Because the green fluorescence rapidly disappears at late M phase, two-dimensional flow cytometry analysis can usually detect a green(high)/red(low) fraction including S-, G(2)- and early M-phase cells but not a transitional fraction between green(high)/red(low) and green(low)/red(low) including late M-phase cells. However, combining the shake-off method concentrated the transitional fraction, which enabled us to separate early and late M-phase cells without using any drugs. Here we demonstrate for the first time that cells in early M phase are more radiosensitive than those in late M phase, implying that early M phase is the most radiosensitive sub-phase during the cell cycle. |
| Databáze: | OpenAIRE |
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