辐射对肿瘤微环境中影响.docxVIP

辐射对肿瘤微环境中Treg细胞及其相关因子的影响邵立虹 金霖霖 左斯尧 魏巍 董卓 张海芹 韩海玲 高辉 金顺子[摘要] 目的 检测照射后与肺腺癌A549细胞共培养的淋巴细胞中调节性T细胞（Treg细 胞）及其相关细胞因子和共刺激分子表达的变化，探讨辐射对肿瘤微环境中免疫耐受机制的影响。 方法 分离人外周血淋巴细胞单独培养或与肺腺癌A549细胞进行共培养一段时间后，以6GyX射 线进行照射，继续共培养一段时间后，以流式细胞术检测淋巴细胞中CD4+ CD25+、CD4+ CD25+ FoxP3+、CD4+ CD25+ NrP1+不同亚组的百分数、共刺激分子ICOS和CTLA-4的表达情况;ELISA法 检测培养体系上清中IL-10、TGF-β、IL-17的分泌量。结果 与照射前后单独培养组相比，相应条件 下共培养组中CD4+ CD25+、CD4+ CD25+ FoxP3+、CD4+ CD25+ NrP1+、ICOS、CTLA-4、IL-10和 TGF-β比例均增加(t=2.78～40.61，P＜0.05)。而与照射前相比，照射后单独培养组和共培养组的 CD4+ CD25+和CTLA-4比例增加(t=2.96、2.94、4.47、2.77，P＜0.05)。细胞因子IL-17的表达量 在肺腺癌A549细胞和X射线分别作用下无明显变化，但共同作用后会明显增加(t=4.00、4.71， P ＜0.05)。结论 辐射刺激肺癌细胞肿瘤微环境中的CD4+ CD25+Treg细胞亚组及其相关 CTLA-4和IL-17的表达，诱导肿瘤微环境中的免疫耐受作用。电离辐射;A549细胞系;肿瘤微环境;Treg细胞;免疫耐受Effects of the microenvironment of irradiated tumor cells on Treg cells and related regulation factors Shao LihongJin LinlinZuoSiyaoWei WeiDongZhuoZhang HaiqinHan HailingCollege of Public Medicine, KeyLaboratory ofRadiobiology,MinistryofHealth,Jilin University,Changchun 130021,China[ Abstract] Objective To explorethe immune tolerance intumor microenvironment, wedetected the expressions of CD receptors and related cytokines in Treg cells after being co-cultured with irradiated lung cancer cells. Methods Lymphocyteswere isolatedfromhumanperipheral blood and culturedwith lung adenocarcinoma cells A549, after irradiation with 6 GyX-rays, theexpression of CD4+CD25+,CD4 + CD25 + FoxP3+ , CD4 + CD25 + NrP1+ and ICOS, CTLA-4 in the lymphocytes were tested by flowcytometry,and the contents of IL-10, TGF-β and IL-17in thesupernatantwere detectedby ELISA. Results Compared with lymphocytes alone culture, theexpressions of CD4+ CD25+, CD4+ CD25 + FoxP3+, CD4 + CD25 + NrP1 +, ICOS, CTLA-4, IL-10 and TGF-β in the co-culture groupswithand without irradiation were allincreased(t=2.78- 40.61,P＜ 0. 05),the CD4+CD25+andCTLA-4in the lymphocytes were increased after irradiation ( t = 2.96,2. 94,4.47,2.77, P ＜ 0. 05 ). The content ofIL-17 inthe supernant were increased afterco-cuturewithirrad