中低剂量X射线照射对肿瘤相关巨噬细胞极化类型改变的影响研究  

Effect of medium and low doses X-ray irradiation on polarization type changes of tumor associated macrophages

作　　者：孙超男 刘羽欣[1] 程云云 吴宇晨 林纪国 王煜垚 赵刚[1] SUN Chaonan;LIU Yuxin;CHENG Yunyun;WU Yuchen;LIN Jiguo;WANG Yuyao;ZHAO Gang(NHC Key Laboratory of Radiobiology,School of Public Health,Jilin University,Changchun 130021,China)

机构地区：[1]吉林大学公共卫生学院,国家卫生健康委员会放射生物学重点实验室,长春130021

出　　处：《中国免疫学杂志》2024年第12期2465-2470,2477,共7页Chinese Journal of Immunology

基　　金：国家自然科学基金青年科学基金项目(8230482)。

摘　　要：目的:探讨中低剂量X射线照射对肿瘤相关巨噬细胞(TAMs)极化类型改变的影响及机制。方法:采用小鼠乳腺癌4T1细胞培养上清(4T1-TCS)培养小鼠RAW264.7巨噬细胞系,诱导其成为M2型TAMs,在此基础上进行中低剂量X线照射,Griess法检测NO,流式细胞术检测CD86和CD206,ELISA检测细胞因子IL-10和IL-12p70,免疫荧光检测iNOS和Arg-1,Western blot检测极化相关通路蛋白,观察中低剂量照射是否可以改变TAMs极化类型。结果:4T1-TCS诱导的TAM NO分泌减少,CD206、Arg-1表达升高,iNOS表达降低;低剂量照射(0.075 Gy、0.5 Gy、1 Gy)TCS-TAM,与未经照射组相比,NO分泌、CD86、CD206、Arg-1、iNOS表达均无显著变化;中等剂量(2 Gy、4 Gy、6 Gy)照射TCS-TAM,NO分泌增多,流式结果显示CD86表达升高,CD206表达减少;免疫荧光结果显示,细胞iNOS表达升高,Arg-1表达减少;ELISA结果显示细胞IL-12p70分泌增多,IL-10分泌减少;Western blot结果显示NOX2、ATMS1981*、IRF5蛋白表达均升高。结论:4T1-TCS可成功诱导巨噬细胞RAW264.7极化为M2表型,低剂量照射对TCS-TAM极化表型无显著影响,中等剂量照射可诱导TCS-TAM极化为抑制肿瘤生长的M1表型,且极化的翻转机制可能与IRF5通路有关。Objective:To explore effects and mechanisms of medium and low doses X-ray irradiation on polarization type changes of tumor associated macrophages(TAMs).Methods:Mouse RAW264.7 macrophage line was cultured with supernatant of 4T1 tumor cell culture(4T1-TCS)of mouse breast cancer,and induced to M2 TAMs.On this basis,medium and low doses X-ray irradiation was conducted,and NO was detected by Griess method,CD86 and CD206 were detected by flow cytometry.ELISA was use to detect cytokines IL-10 and IL-12p70,immunofluorescence was use to detect iNOS and Arg-1.Western blot was used to detect polarization related pathway proteins,and whether medium and low doses of radiation could alter polarization type of TAMs was observed.Results:4T1-TCS induced TAM had decreasing NO secretion and increasing CD206 and Arg-1 expressions,iNOS expression was decreased;with low doses irradiation(0.075 Gy,0.5 Gy,1 Gy)of TCS-TAM,compared with non irradiation group,NO secretion,expressions of CD86,CD206,Arg-1 and iNOS showed no significant changes;medium doses(2 Gy,4 Gy,6 Gy)of irradiation on TCS-TAM resulted in increased secretion of NO,and flow cytometry results showed increased expressions of CD86,decreased CD206 expression;immunofluorescence results showed an increase in iNOS expression and a decrease in Arg-1 expression in cells;ELISA results showed an increase in secretion of IL-12p70 by cells and a decrease in secretion of IL-10.Western blot results showed that NOX2,ATMS1981*,IRF5 proteins expressions were increased.Conclusion:4T1-TCS can successfully induce macrophage RAW264.7 polarize into M2 phenotype,while low dose irradiation has no significant effect on polarization phenotype of TCS-TAM.Medium dose irradiation can induce TCS-TAM polarization into M1 phenotype that inhibits tumor growth,and reversal mechanism of polarization may be related to IRF5 pathway.

关 键 词：X射线 肿瘤相关巨噬细胞 极化 通路 

分 类 号：R739.41[医药卫生—肿瘤]