Establishment and identification of radioresistant nasopharyngeal carcinoma cell line
10.3760/cma.j.cn113030-20230410-00071
- VernacularTitle:鼻咽癌放射抵抗细胞株的建立与鉴定
- Author:
Zhenyu YAN
1
;
Xiang CAO
;
Yizhi GE
;
Dan ZONG
;
Xia HE
Author Information
1. 南京医科大学附属肿瘤医院，江苏省肿瘤医院，江苏省肿瘤防治研究所，南京 210009
- Keywords:
Nasopharyngeal carcinoma;
Radiation resistance;
Radioresistance
- From:
Chinese Journal of Radiation Oncology
2023;32(12):1085-1092
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a radioresistant nasopharyngeal carcinoma cell line in vitro and provide experimental basis for further research of the molecular mechanism of radioresistance. Methods:A radioresistant cell line 5-8F-IR was established by dose gradient irradiation. Cell morphological changes were observed by optical microscope. The formation of colony was detected by colony formation assay. Cell viability was detected by CCK-8 assay. Cell proliferation was assessed by 5-ethynyl-2'-deoxyuridine (EdU) assay. DNA damage repair ability was measured by immunofluorescence staining and comet assay. Cell apoptosis and cycle were detected by flow cytometry. The expression levels of DNA damage related protein γH2AX and apoptosis related protein Caspase-3 were measured by Western blot.Results:The 5-8F-IR cells became longer than that of parental generation 5-8F cells after dose gradient irradiation. The colony formation ability ( P<0.01), cell viability ( P<0.001), cell proliferation ability ( P<0.05) and DNA damage repair ability ( P<0.05) of 5-8F-IR cells were significantly stronger than those of parental generation 5-8F cells. The apoptosis rate of 5-8F-IR cells after irradiation was significantly lower than that of parental generation 5-8F cells ( P<0.01). The 5-8F-IR cells showed higher percentage of cells in S phase without irradiation, and obvious G 2/M phase arrest after irradiation ( P<0.01). Western blot showed that the expression levels of γH2AX ( P<0.001) and Caspase-3 ( P<0.05) in 5-8F-IR cells after irradiation were significantly lower compared with those of parental generation 5-8F cells. Conclusion:5-8F-IR cells induced by dose gradient irradiation of human nasopharyngeal carcinoma cell line 5-8F cells exhibit radioresistance and exhibit different biological characteristics compared to their parental 5-8F cells, providing a research tool for exploring radioresistance mechanism of nasopharyngeal carcinoma.
