X线照射增加肺癌细胞A549产生IL-8的研究

X-ray irradiation increases production of IL-8 in lung cancer cell line A549

目的观察照射对肺癌细胞A549产生IL-8的影响及探索其可能机制。方法采用不同剂量X线照射A549细胞,于照射后不同时间收集细胞上清,细胞RNA以及蛋白质,采用RT-PCR检测照射后A549细胞IL-8 mRNA表达水平,并进一步行实时定量PCR验证照射后A549细胞IL-8 mRNA表达水平,行ELISA检测照射后上清中IL-8表达水平,Western Blot检测照射后A549细胞信号通路分子表达情况,采用p38 MAPK抑制剂、NF-κB抑制剂及ROS清除剂预处理细胞,ELISA验证抑制剂对照射诱导A549细胞产生IL-8表达水平的影响。结果照射增加A549细胞的IL-8的表达水平,并具有剂量、时间效应。照射激活A549细胞中p38 MAPK和NF-κB信号通路分子。抑制p38 MAPK和抑制NF-κB能够阻断照射诱导A549细胞产生的IL-8。抑制ROS后并不能抑制照射诱导A549细胞产生的IL-8。结论X线照射能增加A549细胞IL-8的产生,可能与通过激活p38 MAPK和NF-κB信号通路相关,并呈ROS非依赖性模式。

Objective To observe the effect of irradiation on the production of IL-8 in lung cancer cell line A549 and explore its possible mechanism.Methods A549 cells irradiated with different doses of X-rays were used to collect cell supernatant,cellular RNA and protein at different time points after irradiation.The expression level of IL-8 mRNA in A549 cells after irradiation was detected by RT-PCR,which was further validated by real-time quantitative PCR.The expression level of IL-8 in the cell supernatant was quantitatively measured by ELISA.The expression levels of cellular signaling pathway molecules in A549 cells after irradiation were detected byWestern Blot.The A549 cells were pretreated with p38 MAPK inhibitor,NF-κB inhibitor and ROS scavenger.The effect of these inhibitors on the expression of IL-8 in A549 cells induced by irradiation was evaluated by ELISA.Results Irradiation up-regulated the expression of IL-8 in A549 cells in a dose-and time-dependent manner.Irradiation activated the p38 MAPK and NF-κB signaling pathway in A549 cells.p38 MAPK and NF-κB inhibitors blocked the induction of IL-8 of A549 cells by irradiation.Inhibition of ROS failed to inhibit the induction of IL-8 of A549 cells by irradiation.Conclusion Irradiation can increase the production of IL-8 in lung cancer cells A549,possibly through the activation of p38 MAPK and NF-κB signaling pathways in a ROS-independent pattern.