Effect of UbcH5a on the radiosensitivity of esophageal carcinoma cells by regulating the ex-pression of DNA damage repair proteins
Objective:To construct a stable transfected esophageal squamous cell carcinoma EC109 cell line overexpressing human ubiquitin cross-linking enzyme UbcH5a gene and investigate the mechanism of UbcH5a affecting the radiosensitivity of esophageal carcinoma.Methods:EC109 cells were divided into blank control group(no treatment),negative control group(transfected with pEGFP-C1 plasmid)and overexpression group(transfected with pEGFP-UbcH5a plasmid).Real-time fluorescence quantitative PCR(RT-qPCR)and western blotting were respectively used to detect the expression of UbcH5a mRNA and protein to verify the transfection efficiency.Clonogenic assay was used to detect the cell surviving fraction after 2 Gy X-ray irradiation(SF2),and immunofluorescence was used to detect DNA damage.The expression of DNA damage repair related proteins(ATM,ATR,p-ATM,p-ATR,Chk1,Chk2 and BRCA1)was detected by western blotting.Results:Compared with the blank control group,the expression levels of UbcH5a mRNA and protein were increased,SF2 was de-creased,and DNA damage foci were increased in the overexpression group(all P＜0.05),while there was no sig-nificant change in the negative control group(P＞0.05).Compared with the negative control group,the expres-sion of ATM,ATR,p-ATM,P-ATR,Chk1,Chk2 and BRCA1 in the overexpression group was significantly down-regulated before and after X-ray irradiation(all P＜0.05).Conclusion:UbcH5a enhances the radiosensitivity of esophageal cancer cells by inhibiting the expression of DNA damage repair related proteins.
万方数据
维普
