Effect of E1A gene on radiosensitivity of human nasopharyngeal carcinoma cells and its possible mechanism
10.3760/cma.j.issn.1004-4221.2017.11.018
- VernacularTitle:E1A基因通过抑制NF-κB信号通路增加鼻咽癌细胞放射敏感性实验研究
- Author:
Huaping XIAO
1
,
2
;
65212 哥伦比亚,美国密苏里大学医学院Ellis FISCHEL肿瘤中心
;
Qing LI
;
Hui XIE
;
Chunyang LUO
;
Yujiang FANG
Author Information
1. 423000郴州,湘南学院附属医院肿瘤科
2. 65212 哥伦比亚,美国密苏里大学医学院Ellis Fischel肿瘤中心
- Keywords:
E1A gene;
NF-κB;
Casein Kinase 2;
Nasopharyngeal Carcinoma Cell;
Radiosensitivity;
Apoptosis
- From:
Chinese Journal of Radiation Oncology
2017;26(11):1327-1331
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of E1A gene on the radiosensitivity of human nasopharyngeal carcinoma cells and its possible mechanism. Methods The E1A gene was transfected into nasopharyngeal carcinoma CNE-2R cells by adenovirus vector. The expression of E1A gene was detected by RT-PCR. Untransfected CNE-2R cells(PBS group)and CNE-2R cells transfected with empty vector Ad-β-gal(Ad-β-gal group)and E1A(Ad-E1A group)were given 0 Gy,2 Gy,4 Gy,6 Gy,8 Gy 6 MV X-ray irradiation. The changes in radiosensitivity of CNE-2R cells were determined by colony-forming assay. Flow cytometry was used to analyze cell apoptosis in each group. The expression of NF-κB, CK2α, Bcl-2, and cleaved caspase-3 was measured by Western blot. Results RT-PCR confirmed that the E1A gene was transfected into CNE-2R cells and stably expressed. The Ad-E1A group had a significantly lower plating efficiency than the PBS group and the Ad-β-gal group(P<0.05). The Ad-E1A group had significantly lower cell survival rate at 2 Gy irradiation than the PBS group and the Ad-β-gal group(0.217 vs. 0.602, P<0.05;0.217 vs. 0.585, P<0.05). The Ad-E1A group had a significantly higher α/β value than the PBS group and the Ad-β-gal group(24.680 vs. 5.268, P<0.05;24.680 vs. 5.132, P<0.05). Flow cytometry results showed that irradiation alone could promote the apoptosis of CNE-2R cells,when combined with E1A gene,the apoptosis rate was significantly increased(P<0.05). Western blot results showed that E1A gene down-regulated the expression of NF-κB/p65,CK2α,and Bcl-2 and up-regulated the expression of cleaved caspase-3. Conclusions E1A gene can enhance the radiosensitivity of nasopharyngeal carcinoma cells by inhibiting the expression of CK2 to block the NF-κB signaling pathway and promoting cell apoptosis.
