Establishment and comparative analysis of three DNA damage models of GC-1 cells
10.3969/j.issn.1001-1978.2021.01.023
- Author:
Si-Qi YANG
1
;
Hai-Xia ZHAO
1
;
Xu YOU
1
;
Qiong-Yan MA
1
;
Yuan YANG
1
;
Yan ZHANG
1
;
Yong YE
1
;
Jie WU
1
;
Ding YUAN
1
;
Chang-Cheng ZHANG
1
Author Information
1. Medical College of China Three Gorges University
- Publication Type:Journal Article
- Keywords:
8-OHdG;
DNA damage;
GC-1 cells;
P-;
Spermatogonia;
yH2AX
- From:
Chinese Pharmacological Bulletin
2021;37(1):142-148
- CountryChina
- Language:Chinese
-
Abstract:
Aim To establish three kinds of DNA damage models of mouse testicular spermatogonia cell line GC-1 cells and analyze their similarities and differences. Methods GC-1 cells were treated with UVB irradiation, D-galactose(D-Gal) or bleomycin (BLM), respectively. Then the expression and localization of 'Y-H2AX were detected by Western blot and immunofluorescence, the expression and localization of 8-OHdG were measured by immunofluorescence, and the expression levels of p-p53 and p21 were measured by Western blot. Results The expression of -y-H2AX in GC-1 cell reached to the peak 4 h after UVB irradiation and 6 h after D-Gal stimulation, whereas -y-H2AX expression gradually increased after BLM stimulation, and the higher the concentration of BLM,the shorter the time to reach the peak. The results of immunofluorescence showed that 8-OHdG expression was observed in the nucleus and cytoplasm of GC-1 cells after UVB irradiation and BLM stimulation, and the longer the culture time, the more the expression in the nucleus. In contrast, the expression of 8-OHdG was observed in the cytoplasm and reached the peak at 6 h in the D-Gal stimulated GC-1 cells. After UVB irradiation, the protein expression levels of p-p53 gradually increased, while p21 protein expression appeared later than that of p-p53; in the D-Gal stimulated GC-1 cells, the protein expression levels of p-p53 reached the peak at 6 h, and p21 protein expression reached the peak at 12 h; after low concentration BLM stimulation, the protein expression levels of p-p53 and p21 continuously increased, and after high concentration BLM stimulation, the protein expression levels of p-p53 and p21 reached its peak at 2 h, then decreased at 4 h. Conclusions Three kinds of DNA damage models of GC-1 cells are successfully established, and the DNA damage in GC-1 cells treated with D-Gal is mild, while the DNA damage in GC-1 cells treated by UVB irradiation and BLM stimulation is more severe.
