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目的:探讨食管癌细胞照射后细胞周期、细胞凋亡及其相关蛋白表达的变化。方法:食管癌细胞株TE-1和TE-13照射2、5、10、15Gy后,应用流式细胞仪分别检测照射后6、24和48h细胞周期和凋亡指数变化、Westernblot检测细胞周期相关蛋白的表达。结果:2、5、10、15Gy照射0~48h,TE-1和TE-13细胞均出现明显剂量依赖性的G2/M期阻滞和解除变化;5~15Gy照射后48h,TE-13细胞在G2/M期阻滞逐渐解除时伴随着细胞凋亡的明显增加,而TE-1细胞凋亡不增加。2株细胞胞浆CHK2-p68磷酸化水平均呈随照射后时间延长而出现时相性变化,但与照射剂量无关;而CHK1、CHK2、CHK1-p345和CDK1蛋白表达均无明显变化。15Gy照射后24h,TE-13细胞胞浆中cyclinB1表达下降而TE1细胞中无明显变化。结论:病理不同分化的食管癌细胞照射后细胞周期、细胞凋亡以及细胞周期相关蛋白表达变化不尽相同。
Objective: To investigate the changes of cell cycle, apoptosis and related protein expression in esophageal cancer cells after irradiation. Methods: The changes of cell cycle and apoptosis index at 6, 24 and 48 hours after irradiation were detected by flow cytometry after esophageal cancer cell lines TE-1 and TE-13 were irradiated with 2, 5, 10 and 15 Gy respectively. The cell cycle was detected by Western blot Protein expression. Results: At doses of 2, 5, 10, 15 Gy for 0 ~ 48h, the G2 / M phase arrest and release were significantly inhibited in TE-1 and TE-13 cells. In the G2 / M phase arrest gradually relieved with a significant increase in apoptosis, while TE-1 cells did not increase apoptosis. The phosphorylation levels of CHK2-p68 in the two cytoplasm showed time-phased changes with the prolongation of irradiation time, but had no correlation with the dose of CHK2-p68. However, the expression of CHK1, CHK2, CHK1-p345 and CDK1 did not change significantly. After 24h of 15Gy irradiation, the expression of cyclinB1 in the cytoplasm of TE-13 cells decreased but there was no significant change in TE1 cells. Conclusion: The changes of cell cycle, apoptosis and expression of cell cycle related proteins in esophageal cancer cells with different pathological differentiation are different.