| Wu Lei,Ma Jianan,Wang Jingxuan,et al.Analyzing differences in responses of nucleolar and nucleoplasmic compartments to radiation-induced DNA damage using the tFucci(CA)5 cell cycle reporter system[J].Chinese Journal of Radiological Medicine and Protection,2026,46(4):336-343 |
| Analyzing differences in responses of nucleolar and nucleoplasmic compartments to radiation-induced DNA damage using the tFucci(CA)5 cell cycle reporter system |
| Received:November 13, 2025 |
| DOI：10.3760/cma.j.cn112271-20251113-00402 |
| KeyWords:tFucci(CA)5|DNA double-strand breaks|γ-H2AX|Nucleolus|Cell cycle reporter system |
| FundProject:国家自然科学基金(82373526) |
| Author Name | Affiliation | E-mail | | Wu Lei | Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China | | | Ma Jianan | Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China | | | Wang Jingxuan | Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China | | | Sun Jinnuo | Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China | | | Mo Yunqi | Graduate Collaborative Training Base of Academy of Military Sciences, Hengyang Medical School, University of South China, Hengyang 421001, China | | | Cai Dan | Graduate Collaborative Training Base of Academy of Military Sciences, Hengyang Medical School, University of South China, Hengyang 421001, China | | | Wang Zhidong | Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China
Graduate Collaborative Training Base of Academy of Military Sciences, Hengyang Medical School, University of South China, Hengyang 421001, China | | | Shen Liping | Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China | shen_2011yan@sina.com |
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| Abstract:: |
| Objective To observe the responses of the nucleus, nucleolar compartment, and nucleoplasmic compartment of cells in different cycle phases to the DNA double-strand breaks (DSBs) induced by ionizing radiation and to analyze their DSB repair kinetics using the tFucci(CA)5 cell cycle reporter system. Methods A HT29 cell cycle reporter model capable of stably expressing tFucci(CA)5 was constructed by co-transfecting the tFucci(CA)5 and transposase expression plasmids and selecting monoclonal cells. The cells were irradiated with 2 Gy of γ-rays. Post-irradiation, γ-H2AX foci, a DSB marker, were detected at different time points using an immunofluorescence technique. Accordingly, the formation and clearance of γ-H2AX foci in the nucleolar and nucleoplasmic compartments were analyzed to explore the damage levels and repair kinetics of DSBs. Results The established HT29 cell cycle reporter model enabled the precise identification and tracking of cells in phases G1, S, and G2/M. The result indicated that to the initial DSBs induced by radiation, the responses of the nucleolar compartment exhibited greater dependence on cell cycle phases (G1 < S < G2, F = 3.31-187.70, P < 0.05), differing from the damage pattern of the nucleoplasmic compartment (G1 < S/G2, F = 13.63-76.60, P < 0.05). The analysis of DSB repair kinetics reveals that compared to the nucleoplasmic compartment, the nucleolar compartment of cells in the middle (S) and late (G2) phases exhibited significantly higher DSB repair efficiency after irradiation (t =2.41-8.55, P < 0.05). Conclusions This study provides a tool for the intuitive analysis of DNA damage in different cell cycle phases. The result reveal that the responses of the nucleolar and nucleoplasmic compartments to DSBs show significant dependence on the cell cycle. Furthermore, compared to the nucleoplasmic compartment, the nucleolar compartment of cells in the S and G2 phases demonstrates significantly higher DSB repair efficiency. |
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