摘要
目的 了解肺癌细胞株及蛙皮素对树突状细胞 (DC)产生及功能的影响。方法 树突状细胞由健康人外周血单个核细胞CD14+,在完全细胞培养液中加入 10 0 0U /mlGM CSF和 10 0 0U /mlIL 4 ,培养 7天获得 ,肺癌细胞株CRL 5 815 ,CRL 5 82 6 ,Bombesin和Bombesin受体拮抗剂加入培养液中 ,AnnxinV检测DC凋亡 ;流式细胞仪检测CD40 ,CD86 ,CD83,CD80 ,HLA DR阳性表达。结果 培养 7～ 10天后的DC前体细胞表达高水平的HLA DRCD80CD86CD83CD40。肺癌及蛙皮素可导致DC前体细胞凋亡 ,而蛙皮素受体拮抗剂可部分保护蛙皮素致DC凋亡的作用。加入肺癌细胞株或蛙皮素与DC共培养时明显抑制上述细胞表型的表达和DC刺激同种异体T细胞的增殖能力 ,当加入蛙皮素受体拮抗剂时 ,DC细胞表达HLA DRCD80CD86CD83CD40明显增加 ,接近DC正常对照组。结论 肺癌细胞株及蛙皮素可导致DC的凋亡 ,抑制树突状细胞的产生和功能。
Objective To understand the effect of non-small cell lung cancer regulated dendritic cell function and generation. Methods CD14+ peripheral blood mononuclear cells (PBMC) obtained from human healthy volunteers. Cultured DC were collected and separately incubated with human lung cancer lines (CRL-5815, CRL-5826), bombesin and antagonist of bombesin in 6-well plates. Apoptosis of dendritic cells was analysed using Annexin V binding assay. Expressions of CD86, CD83, CD80, CD40, HLA-DR of DC were analyzed by FACscan. Ability of DC to stimulate proliferation of T cells was assessed with allogeneic mixed lymphocyte reactions (MLR) assay. Results Inhibition of DC generation by lung cancer: Dendritic cells from PBMC cultured with GM-CSF 1000U /ml and IL-4 1000U /ml for 7days expressed high levels of CD86, CD83, CD80, CD40, HLA-DR. Dendritic cells co-incubation with lung cancer cells and bombesin inhibited definitely expression of CD86, CD83, CD80, CD40, HLA-DR and significantly decreased ability to induce T cell proliferation (MLR). Antagonist of bombesin blockaded the function of lung cancer-induced apoptosis and generation of DC. Conclusion Non-small cell lung cancer (CRL-5815, CRL-5826) and bombesin inhibited dendritic cell maturation and function and induced apoptosis in vitro.
出处
《肿瘤》
CAS
CSCD
北大核心
2003年第2期115-118,共4页
Tumor
基金
上海市医学学科带头人"百人计划" [31199736 30 2 10 0 0 3]
关键词
肺癌
蛙皮素
树突状细胞
DC
Lung cancer
Bombesin
Dendritic cells(DC)
