| Zhao Wenna,Chen Zihan,Chen Fuqiang,et al.lncRNA ANK3DT affects radiosensitivity of cervical cancer cells by regulating CtIP transcription[J].Chinese Journal of Radiological Medicine and Protection,2025,45(3):170-177 |
| lncRNA ANK3DT affects radiosensitivity of cervical cancer cells by regulating CtIP transcription |
| Received:April 16, 2024 |
| DOI：10.3760/cma.j.cn112271-20240416-00137 |
| KeyWords:Long noncoding RNA Radiosensitivity DNA damage Homologous recombination repair |
| FundProject:国家自然科学基金(81903256) |
| Author Name | Affiliation | E-mail | | Zhao Wenna | Department of Radiation Medicine, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou 510515, China | | | Chen Zihan | Department of Radiation Medicine, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou 510515, China | | | Chen Fuqiang | Department of Radiation Medicine, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou 510515, China | | | Du Jie | Department of Radiation Medicine, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou 510515, China | | | Zhou Meijuan | Department of Radiation Medicine, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou 510515, China | lkzmj@smu.edu.cn |
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| Abstract:: |
| Objective To explore the effect of long noncoding RNA ANK3DT (lncRNA ANK3DT) on DNA damage repair ability and radiosensitivity of cervical cancer HeLa cells. Methods The effect of lncRNA ANK3DT on the repair efficiency of DNA double-strand breaks (DSBs) was analyzed by the pre-constructed quantitative assay system based on the CRISPR/Cas9 system targeting homologous recombination (HR) and non-homologous end-joining (NHEJ) repair. Clone formation assay, flow cytometry, and immunofluorescence were used to detect the effects of down-regulation of lncRNA ANK3DT on cellular radiosensitivity, apoptosis and G2/M phase arrest, and DSB repair after X-ray irradiation. Western blot, qPCR, and dual fluorokinase reporter gene plasmid was used to detect the effects of lncRNA ANK3DT on the HR repair-related protein CtIP expression and transcriptional regulation. Results Down-regulation of lncRNA ANK3DT significantly inhibited HR repair and had no effect on NHEJ repair. The expression of lncRNA ANK3DT was significantly increased in HeLa cells at 24, 48, 72 h after X-ray irradiation (t= -23.39, -88.83, -52.42,P< 0.05). Down-regulation of lncRNA ANK3DT inhibited X-ray-induced DSB repair and increased apoptosis levels (t= -14.63, P< 0.05), prolonged G2/M phase block (t= -19.50, P< 0.05), increased the radiosensitivity of HeLa cells (radiosensitization ratio=1.21), inhibited the CtIP promoter activity and decreased its mRNA and protein expression. Conclusions lncRNA ANK3DT affects HR repair by regulating CtIP transcription, and down-regulation of lncRNA ANK3DT increases the radiosensitivity of HeLa cells by inhibiting DSB repair after ionizing radiation. |
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