龙金华,曾宪琳,金仙槐
LONG Jinhua,ZENG Xianlin,JIN Xianhuai

• 1:贵州医科大学附属肿瘤医院头颈肿瘤科
• 2:贵州医科大学生物与工程学院

摘要(Abstract)：

目的 研究大分割电离辐射对鼻咽癌(NPC)细胞与树突状细胞(DCs)共培养后DCs免疫表型和迁移能力的影响。方法 核磁共振检测放化疗后NPC患者(NPC组)鼻咽部肿瘤变化，流式细胞术检测NPC组及健康志愿者(健康对照组)外周血DCs数量分群[髓系DCs(CD11c~+ myeloid DCs, MDCs)和浆系DCs(CD123~+ plasmacytoid DCs, PDCs)]情况；将DCs细胞分为未成熟树突状细胞组(imDCs组)、成熟DCs组(mDCs组)、imDCs+4 Gy组以及imDCs+18 Gy组，后两组是将imDCs与大分割电离辐射(4 Gy、18 Gy)照射后的NPC细胞共培养所得；流式细胞术、Transwell小室实验以及蛋白免疫印迹(Western blot)分别检测各组DCs免疫表型[人白细胞抗原DR(HLA-DR)、白细胞分化抗原(CD80)、CD83、CD86、CC基序趋化因子受体7(CCR7)]、细胞迁移能力以及细胞中环磷酸鸟苷-磷酸腺苷合成酶(cGAS)和干扰素基因刺激因子(STING)蛋白的表达。结果 与放疗前相比，放疗后NPC组患者鼻咽黏膜稍增厚，肿瘤明显退缩，外周血中MDCs的数量明显减少(P<0.001);与imDCs组比较，imDCs+4 Gy组以及imDCs+18 Gy组中imDCs的免疫表型分子HLA-DR、CD80、CD83、CD86及CCR7的表达显著上调(P<0.001),且GAS和STING蛋白的表达增加(P<0.05);Transwell实验结果显示，DCs迁移能力随着电离辐射剂量的增加而增强(P<0.05)。结论 经大分割电离辐射照射的NPC细胞可诱导imDCs向mDCs分化，这可能产生特异性的抗肿瘤免疫应答。
Objective To investigate the effect of hypofractionated ionizing radiation on the immunophenotype and migration capacity of dendritic cells(DCs) after co-culturing with nasopharyngeal carcinoma(NPC) cells. Methods Nuclear magnetic resonance imaging was used to detect the changes in nasopharyngeal carcinoma in NPC patients(NPC group) after radiotherapy and chemotherapy. Flow cytometry was used to detect peripheral blood DCs populations in NPC group and healthy volunteers(healthy control group). DCs cells were divided into immature dendritic cells group(imDCs group), mature DCs group(mDCs group), imDCs+4 Gy group, and imDCs+18 Gy group. The latter two groups were obtained by co culturing imDCs with irradiated NPC cells using 4 Gy and 18 Gy, respectively. Flow cytometry, Transwell assay and Western blot were used to detect the immune phenotype(HLA-DR, CD80,CD83,CD86,CCR7),cell migration capacity and the protein expressions of cyclic guanosine phosphate adenosine synthetase(cGAS) and interferon gene stimulating factor(STING) in DCs of each group. Results When compared with before radiotherapy, NPC group exhibited slight thickening of the nasopharyngeal mucosa, significant tumour regression and a significant decrease in mDC number in peripheral blood(P<0.001). When compared with imDCs group, the expression levels of immunophenotypic molecules HLA-DR, CD80, CD83, CD86, and CCR7 in imDCs+4 Gy group and imDCs+18 Gy group were remarkably upregulated(P<0.001), The protein expression levels of GAS and STING were also upregulated(P<0.05). Transwell results showed that the migration capacity of DCs was increased with the increase of ionizing radiation dose(P<0.05). Conclusion Hypofractionated ionizing radiated-NPC cells may induce imDCs to differentiate into mDCs, which may trigger a specific anti-tumour immune response.

关键词(KeyWords)： 大分割电离辐射;鼻咽癌;细胞培养;树突状细胞;免疫表型;环磷酸鸟苷-磷酸腺苷合成酶;干扰素基因刺激因子
hypofractionated ionizing radiation;nasopharyngeal carcinoma;cell culture;dendritic cells;immunophenotype;cyclic guanosine phosphate adenosine synthetase;interferon gene stimulating factor

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目（82060555）

作者(Author): 龙金华,曾宪琳,金仙槐
LONG Jinhua,ZENG Xianlin,JIN Xianhuai

DOI: 10.19367/j.cnki.2096-8388.2023.09.008

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