lncRNA ANK3DT affects radiosensitivity of cervical cancer cells by regulating CtIP transcription
10.3760/cma.j.cn112271-20240416-00137
- VernacularTitle:lncRNA ANK3DT调控CtIP转录影响宫颈癌细胞的放射敏感性
- Author:
Wenna ZHAO
1
;
Zihan CHEN
1
;
Fuqiang CHEN
1
;
Jie DU
1
;
Meijuan ZHOU
1
Author Information
1. 南方医科大学公共卫生学院放射医学系 广东省热带病研究重点实验室，广州 510515
- Publication Type:Journal Article
- Keywords:
Long noncoding RNA;
Radiosensitivity;
DNA damage;
Homologous recombination repair
- From:
Chinese Journal of Radiological Medicine and Protection
2025;45(3):170-177
- CountryChina
- Language:Chinese
-
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 G 2/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 G 2/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.
