Effects of silencing single-stranded DNA-binding protein 1 gene on proliferation and DNA repair of submandibular gland cells after irradiation
10.3760/cma.j.issn.0254-5098.2017.09.001
- VernacularTitle:单链DNA结合蛋白1沉默对颌下腺细胞放射后增殖修复的影响
- Author:
Long CHEN
1
;
Qiuli LYU
;
Binling YI
;
Zhe SUN
;
Daiyou WANG
Author Information
1. 广西医科大学附属口腔医院口腔颌面外科 广西口腔颌面修复与重建研究重点实验室 广西颌面外科疾病诊治研究高校重点实验室
- Keywords:
Single-stranded DNA-binding protein 1;
Submandibular gland cell;
Ionizing radiation;
Double-stranded DNA breaks
- From:
Chinese Journal of Radiological Medicine and Protection
2017;37(9):645-650
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of silencing the gene of single-stranded DNA-binding protein 1 (SSB1) on proliferation and DNA repair of rat submandibular gland (SMG) cells after irradiation, and explore the relationship between SSB1 and DNA damage repair. Methods Primary rat SMG cells were obtained by mechanical-enzyme digestion and identified by immunohistochemistry. The cells were divided into three groups, including blank control, negative control and shRNA transfection group. The shRNA was transfected into cells by recombinant adenovirus vector. Real-time quantitative PCR ( qRT-PCR) was used to detect the expression of SSB1 after silencing. The cell viability was detected by CCK-8 assay. Immunofluorescence analysis was performed to observe the dynamic formation of γ-H2AX foci. Results The SMG cells were positively stained for both Pan CK and α-Amylase. The efficiency of shRNA transfection was about 90%at 72 h post-transfection. Compared with the blank control group, the expression of SSB1 was significantly decreased in the cells transfected with shRNA (t=16. 24, P<0. 05). The cell viability of shRNA transfection group without irradiation was decreased indistinctively and became lower than the blank control group significantly until 120 h(t=3. 29, P<0. 05). After radiation with 5 Gy of γ-rays, the cell viability of shRNA transfection group was lower than that of the control groups significantly (F=10. 19-30. 13, P<0. 05). Silencing the expression of SSB1 could increase the number ofγ-H2AX foci in SMG cells at different time of radiation. Conclusions After silencing of the expression of SSB1, the SMG cells could be more radiosensitive, which indicats that SSB1 may play an important role in DNA damage repair after radiation.
