电离辐射对脑胶质瘤cGAS-STING-IFNⅠ信号通路的调控作用及其机制
Radiation on the regulation of Glioma cGAS-STING- IFNⅠ pathway and its mechanism
环磷酸鸟苷-腺苷合成酶（cyclic guanosine monophosphate-adenosine monophosphate synthase, cGAS）是一种胞质DNA感受器，同过识别胞质中的双链DNA(double strand DNA, dsDNA)，催化腺苷三磷酸（adenosine triphosphate, ATP）和鸟苷三磷酸（guanosine triphosphate, GTP）形成环鸟苷酸腺苷酸（cyclic guanosine monophosphate-adenosine monophosphate, cGAMP）。cGAMP激活干扰素基因刺激分子（stimulator of interferon gene, STING），进而产生Ⅰ型干扰素（Type I interferon，IFNⅠ）及其他细胞因子。目的：探究电离辐射对脑胶质瘤cGAS-STING-IFNⅠ信号通路的调控作用及其机制。方法：不同剂量X射线照射人胶质瘤细胞，通过免疫荧光染色检测dsDNA在胞浆中的含量；通过实时定量PCR检测照后cGAS、干扰素刺激基因（interferon stimulating genes，ISGs）（如MX肌动蛋白样GTP酶1（MX dynamin like GTPase 1，MX1）、C-X-C基序趋化因子10（C-X-C motif chemokine ligand 10，CXCL10）、β1干扰素（interferon beta 1，IFNβ1）等）和三重修复核酸外切酶1（three prime repair exonuclease 1，TREX1）基因的相对表达量；利用Western blot 检测照后细胞内干扰素调节因子3（interferon regulatory factor 3, IRF3）、p-IRF3蛋白表达量的变化；通过ELISA实验检测照后细胞内2\'3\'-cGAMP和上清中β干扰素（interferon-beta，IFN-β）的产量。结果：照后常氧脑胶质瘤细胞胞浆中的dsDNA的累积量，cGAS和ISGs（MX1、IFNβ1、CXCL10）mRNA的相对表达量，p-IRF3蛋白表达量，细胞中2\'3\'-cGAMP和上清液中IFN-β的产量，均在0-16 Gy时随着照射剂量的增加而增加，而在单次大剂量辐射（24 Gy）中反而减少。乏氧明显降低辐射引起的细胞胞浆中dsDNA的累积量，cGAS和ISGs的相对表达量，p-IRF3的蛋白表达量，细胞中2\'3\'-cGAMP和上清液中IFN-β的产量。结论：电离辐射对脑胶质瘤cGAS-STING-IFNⅠ信号通路的激活在0-16 Gy范围内随着照射剂量的增加而增强，而乏氧可明显抑制电离辐射对该通路的激活作用。
he cGAS, is a cytoplasmic DNA susceptor, which identifies the dsDNA in the cytoplasm and catalyzes ATP and GTP to form cGAMP. cGAMP activates STING, thereby produces type I interferon and other cytokines. Objective: To explore the regulation of radiation on cGAS-STING-IFNI pathway in glioma and its mechanism. Method: The distribution of dsDNA in cytoplasm is detected by Immunofluorescence staining; The relative expression of cGAS, ISGs (MX1, CXCL10, IFNβ1) and Trex1 are detected by real-time quantitative PCR; Changes in the expression of IFR3 and p-IRF3 protein were detected by Western blot; cGAMP and IFN-β were detected by ELISA experiments. Results: Under normoxic condition, the accumulation of dsDNA in the cytoplasm of glioma cells, the relative expression of cGAS and ISGs, the expression of p-IRF3 protein, and the production of 2\'3\'-cGAMP and IFN-β in the supernatant increased with the increase of radiation dose in 0-16 Gy, but decreased in single high dose radiation (24 Gy). Hypoxia reduced the accumulation of dsDNA in the cytoplasm, the relative expression of cGAS and ISGs, the protein expression of p-IRF3, and the production of 2\'3\'-cGAMP and IFN-β in the supernatant. Conclusion: The activation of cGAS-STING-IFNI pathway by radiation is dose-dependent in 0-16 Gy, which could be inhibited by hypoxia in glioma cells.
黄文鹏、修増赫、薛雪菲、何玉瓶、杨巍、杨颖、杨双玉
基础医学肿瘤学神经病学、精神病学
电离辐射cGAS-STING- IFNⅠ乏氧脑胶质瘤
Ionizing radiationcGAS-STING-IFNIhypoxiaglioma
黄文鹏,修増赫,薛雪菲,何玉瓶,杨巍,杨颖,杨双玉.电离辐射对脑胶质瘤cGAS-STING-IFNⅠ信号通路的调控作用及其机制[EB/OL].(2022-03-22)[2025-07-03].http://www.paper.edu.cn/releasepaper/content/202203-319.点此复制
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