Effects of LEF1 and CTNNB1 on cycle arrest, apoptosis and radiation resistance of esophageal carcinoma cells
Ge Dahe1, Liu Yaqing1, Nan Jun1, Li Jian2, Ding Hui2, QiaoShulin3, An Yuan4, Xu Chunjin1
1Department of Gastroenterology,Shangqiu First People's Hospital,Shangqiu 476100,China; 2Department of Gastroenterology,Henan Provincial People's Hospital,Zhengzhou 450003,China; 3Department of Oncology and Radiotherapy,Shangqiu First People's Hospital,Shangqiu 476100,China; 4Outpatient Department ofChina Institute of Water Resources and Hydropower Research,Beijing 100038,China
Abstract：Objective To investigate the relationship between the expression level of lymphocyte enhancer-binding factor 1(LEF1) and CTNNB1 and the cycle arrest, apoptosis and radiation resistance of esophageal cancer cells and unravel the related mechanisms. Methods Recombinantplasmids and empty plasmids expressing LEF1 and CTNNB1were constructed and transfected into esophageal cancer cells. RT-PCR assay was used to detect the transfection efficiency of the plasmids. Clone formation assay, CCK8 assay, cell cycle test by flow cytometry, apoptosis test by flow cytometry and Western blot were performed to detect the differences in theradioresistance, proliferation, cell cycle and apoptosis of esophageal cancer cells before and after transfection. Results The survival rate of clonal colony cells in the pGEX-LEF1+pCMV6-CTNNB1 group was significantly better than those in other groups (P<0.05). The proliferation of clonal colony cellsat 72h, 96h and 120h in the pGEX-LEF1+pCMV6-CTNNB1 group was significantly better than those in the pGEX+pCMV6,pGEX-LEF1+pCMV6 and pCMV6-CTNNB1+pGEX groups (all P<0.05). The percentage of G2 phase arrest cells in the pGEX-LEF1+pCMV6-CTNNB1 group was significantly higher than those in the other groups (all P<0.05). The apoptosis rate of esophageal cancer cells in the pGEX-LEF1+pCMV6-CTNNB1 group was significantly lower compared with those in the pGEX+pCMV6, pGEX-LEF1+pCMV6 and pCMV6-CTNNB1+pGEX groups (all P<0.05). The expression levels of Bax and Caspase 3 proteins in the pGEX-LEF1+pCMV6-CTNNB1 group were significantly lower than those in the pGEX+pCMV6, pGEX-LEF1+pCMV6 and pCMV6-CTNNB1+pGEX groups (all P<0.05). The expression level of Bcl-2 protein in the pGEX-LEF1+pCMV6-CTNNB1 group was significantly higher compared with those in the other groups (all P<0.05). Conclusion LEF1 and CTNNB1 can regulate the proliferation and G2 phase arrest of esophageal cancer cells after radiation intervention by mediating the Wnt signaling pathway, and improve the radiation resistance of esophageal cancer cells by inhibiting cell apoptosis.
Ge Dahe,Liu Yaqing,Nan Jun et al. Effects of LEF1 and CTNNB1 on cycle arrest, apoptosis and radiation resistance of esophageal carcinoma cells[J]. Chinese Journal of Radiation Oncology, 2022, 31(4): 376-382.
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