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X射线对A549细胞DNA的损伤可能与JAK/STAT信号通路的激活有关

彭丽桥¹, 李程豪², 毛兵^1,*

¹四川大学华西医院中西医结合科，成都 610000；²甘肃中医药大学甘肃省高校重大疾病分子医学与中医药防治研究省级重点实验室，兰州 730000

摘要

本研究旨在观察不同剂量X射线对A549细胞DNA的损伤和JAK/STAT信号通路激活水平之间的关系。分别用2、4、8 Gy X射线对A549细胞进行照射后，用CCK8法检测A549细胞增殖情况，用酶联免疫法检测照射后不同时间点培养液上清中白介素6 (interleukin 6, IL-6)的含量，用免疫荧光染色法检测细胞IL-6受体(IL-6 receptor, IL-6R)和p53结合蛋白1 (p53 binding protein 1, 53BP1)的蛋白表达情况，用Western blot检测细胞JAK2、p-JAK2、STAT3和p-STAT3的蛋白表达水平。结果显示，和对照组相比，X射线照射可降低细胞增殖水平，上调53BP1表达，提高细胞培养液上清中IL-6含量，并上调IL-6R、JAK2、p-JAK2、STAT3和p-STAT3表达水平。X射线照射的上述作用存在一定的剂量依赖性。以上结果提示，X射线造成细胞DNA损伤的机制可能与JAK/STAT信号通路的激活有关。

关键词： X射线; A549细胞; JAK/STAT信号通路

分类号：R734.2

Activation of the JAK/STAT signal pathway may be involved in DNA damage of A549 cells induced by X-ray

PENG Li-Qiao¹, LI Cheng-Hao², MAO Bing^1,*

¹Department of Integrated Traditional Chinese and Western Medicine, West China Hospital, Sichuan University, Chengdu 610000, China；²Provincial-level Key Lab for Molecular Medicine of Major Diseases and the Prevention and Treatment with Traditional Chinese Medicine Research in Gansu Colleges and Universities, Gansu University of Chinese Medicine, Lanzhou 730000, China

Abstract

The aim of this study was to investigate the relationship between the effects of different doses of X-rays on DNA damage and JAK/STAT signaling pathway activation in A549 cells. The A549 cells were radiated with X-rays at doses of 2, 4, and 8 Gy. The proliferation of A549 cells was detected by CCK8 method. The content of interleukin 6 (IL-6) in culture medium at different time points after irradiation was detected by enzyme-linked immunoassay, and the expression levels of IL-6 receptor (IL-6R) and p53 binding protein 1 (53BP1) were detected by immunofluorescent staining. The expression levels of JAK2, p-JAK2, STAT3 and p-STAT3 were detected by Western blot. The results showed that, compared with the control group, X-ray irradiation reduced the cellular proliferation, up-regulated the expression of 53BP1, increased the IL-6 content in the medium supernatant, and up-regulated the protein expression levels of IL-6R, JAK2, p-JAK2, STAT3, and p-STAT3. The above effects of X-ray irradiation were dose-dependent. These results suggest that the mechanism by which X-rays cause DNA damage in A549 cells may involve activation of the JAK/STAT signaling pathway.

Key words: X-ray; A549 cells; JAK/STAT signal pathway

收稿日期：2018-12-13　　录用日期：2019-06-02

通讯作者：毛兵　　E-mail: maobing@medmail.com.cn

DOI: 10.13294/j.aps.2019.0068

引用本文：

彭丽桥, 李程豪, 毛兵. X射线对A549细胞DNA的损伤可能与JAK/STAT信号通路的激活有关[J]. 生理学报 2019; 71 (5): 698-704.

PENG Li-Qiao, LI Cheng-Hao, MAO Bing. Activation of the JAK/STAT signal pathway may be involved in DNA damage of A549 cells induced by X-ray. Acta Physiol Sin 2019; 71 (5): 698-704 (in Chinese with English abstract).