食管癌细胞系TE13R120放射抗性与HDAC3、NF-kB和NRAGE关系的研究

目的探讨HDAC3、NF—kB和NRAGE基因与食管癌细胞放射抗性的关系。方法以人食管癌细胞系TE13和由TE13经γ射线反复照射建立的放射抗性细胞系TE13R120为研究对象，用Western blotting技术检测这两种细胞于不同时间、剂量点照射后基因HDAC3、NF—kB和NRAGE的蛋白表达；用流式细胞术检测相同时间、剂量点两种细胞的凋亡和细胞周期分布情况。结果Western blotting显示，与TE13相比，TE13R120细胞照射前后HDAC3的核表达均增高，NF-kB的表达也明显增强，NRAGE在两种细胞胞浆中的表达无差异。与TE13相比，TE13R120中S期细胞明显增多，照射后G2期阻滞明显，凋亡水平较低。结论基因HDAC3和NF-kB可能在食管癌细胞放射抗性形成中发挥作用并可能有协同作用；NRAGE与食管癌辐射抗性的关系有待进一步研究。

A study of the relationship of the radioresistance of esophageal carcinoma cell line TE13R120 with HDAC3, NF-kB and NRAGE

Objective To evaluate the relationship of the radioresistance of esophageal carcinoma with genes HDAC3, NF-kB and NRAGE. Methods Two cell lines are chosen for the study, esophageal carcinoma cell line TE13 and esophageal carcinomatous radioresistant cell line TE13R120 which is derived from TE13 by fractionated ionizing radiation. The protein expression of genes HDAC3, NF-kB and NRAGE were measured by Western blotting in the two cell lines at different time intervals and 12 h after different radiating dosages. Apoptotic cell population and cell cycle distribution of the two cell lines were detected by FCM through the same time and dosage-courses with above. Results The protein expressions of NF-kB and HDAC3 in TE13R120 cells were elevated before and after radiation compared with those of TE13 cells. No change of NRAGE expression had been found in cytoplasma of TE13R120 cells compared with TE13 cells. The percentages of TE13R120 cells in G 2 phase were increased at 4, 16 and 24 h after 4 Gy radiation and at 12 h after 2, 6 and 10 Gy radiation. In contrast, no significant change had been observed in TE13 in G 2 phage after radiation. Additionally, the percentages of TE13R120 cells in S phage were significantly higher than those of TE13 cells before and 12 h after different doses radiation. The apoptosis percentages of TE13R120 cell is slightly higher than those of TE13. Conclusion The increasing protein expression of HDAC3 and NF-kB in TE13R120 suggest a very important and possibly, a cooperative effect of these two genes to induce the radioresistance of TE13R120 cell. Further study should be done to reveal the relationship of NRAGE with the radioresistance of esophageal carcinoma.