摘要
目的探讨干扰素基因激活因子(STING)激动剂对皮肤黑色素瘤细胞的放射增敏作用及其机制。方法为检测STING激动剂环二腺苷酸(c-di-AMP)对人皮肤黑色素瘤细胞(A375)辐射后的影响,将人皮肤黑色素瘤细胞A375分为空白对照组、10μmol/L c-di-AMP处理组、X射线照射组、照射+c-di-AMP组。通过CCK-8法、乳酸脱氢酶(LDH)释放实验、平板克隆形成实验以及流式细胞术等手段,检测c-di-AMP对A375细胞的放射增敏作用。采用Western blot检测细胞死亡相关蛋白表达。结果10μmol/L的c-di-AMP联合10 Gy X射线照射时可见明显的放射增敏效应,A375细胞活力较X射线照射组显著下降(t=5.11,P<0.05),细胞毒性显著增加(t=10.15,P<0.05),细胞凋亡显著增加(t=4.41,P<0.05),细胞克隆增殖能力显著降低(t=6.30、3.55、5.45、3.55,P<0.05)。c-di-AMP的放射增敏比(SER)为1.88。10μmol/L的c-di-AMP联合10 Gy X射线后细胞死亡相关信号通路(细胞凋亡、细胞坏死、铁死亡)相关蛋白表达较X射线照射组显著上调。结论采用c-di-AMP激活STING可显著增加电离辐射对皮肤黑色素瘤的放射敏感性,为皮肤黑色素瘤放射治疗提供了新的策略。
Objective To investigate the radiosensitizing effect and underlying mechanism of STING agonist(c-di-AMP)on cutaneous melanoma cells.Methods Human cutaneous melanoma cells(A375)were divided into four groups:the control group,10μmol/L c-di-AMP group,X-ray irradiation group and X-ray irradiation combined with c-di-AMP group.The radiosensitizing effect of c-di-AMP on A375 cells was detected by CCK-8-based viability assay,lactate dehydrogenase(LDH)release assay,flow cytometry-based apoptosis assay,and colony formation assay.Western blot analysis was used to determine the expressions of cell death-related proteins.Results In combination with 10 Gy X-ray irradiation,10μmol/L c-di-AMP showed significant radiosensitization effect in A375 cells,which was evidenced by decreased cell activity(t=5.11,P<0.05),increased cytotoxicity(t=10.15,P<0.05)and cell apoptosis(t=4.41,P<0.05)and reduced clone viability(t=6.30,3.55,5.45,3.55,P<0.05).The calculated radiosensitization ratio of c-di-AMP to A375 cells was 1.88.Moreover,10μmol/L c-di-AMP further increased the expressions of cell death-related proteins induced by radiation in A375 cells.Conclusions The STING agonist c-di-AMP can be used as a radiosensitizer for cutaneous melanoma,which may provide a novel strategy for radiotherapy.
作者
唐绍凯
陈建辉
耿凤豪
杨婷仪
李晓倩
杜小波
张舒羽
Tang Shaokai;Chen Jianhui;Geng Fenghao;Yang Tingyi;Li Xiaoqian;Du Xiaobo;Zhang Shuyu(Laboratory of Radiation Medicine,West China School of Basic Medical Sciences&Forensic Medicine,Sichuan University,Chengdu 610041,China;NHC Key Laboratory of Nuclear Technology Medical Transformation,Mianyang Central Hospital,Mianyang 621099,China)
出处
《中华放射医学与防护杂志》
CAS
CSCD
北大核心
2023年第1期1-7,共7页
Chinese Journal of Radiological Medicine and Protection
基金
国家自然科学基金(82073477)
四川省杰出青年科技人才项目(2022JDJQ0051)
四川大学-泸州科技创新研发项目(2021CDLZ-9)。
