非小细胞肺癌中血管内皮生长因子对树突状细胞的抑制作用

Vascular endothelial growth factor inhibits dendritic cells from patients with non-small cell lung carcinoma

目的 探讨非小细胞肺癌 (NSCLC)中血管内皮生长因子 (VEGF)对树突状细胞 (DC)的影响。方法 用流式细胞仪对 85例NSCLC患者 (NSCLC组 )及 14名正常人 (正常人组 )外周血DC含量进行检测 ,并用酶联免疫吸附 (ELISA)法检测血浆VEGF165浓度。体外条件下由正常人 (血站献血者 )外周血CD+ 14 单个核细胞 (PBMC)培养DC。实验分 2组 ,对照组于培养第 1天和第 4天分别加入rhGM CSF 10 0 0U/ml及rhIL 4 80ng/ml;VEGF165实验组在此基础上于培养第 1天及第 4天加入rhVEGF1655 0ng/ml,以验证其对DC分化和生存的影响 ,用流式细胞仪分析培养细胞的细胞表型及凋亡率。结果 NSCLC组患者血浆VEGF165浓度明显高于正常人组 (P <0 0 5 ) ,而外周血DC含量明显低于正常人组 (P <0 0 1)。NSCLC组患者血浆VEGF165浓度与外周血DC含量呈负相关 (P <0 0 5 ) ,两者与患者的性别、年龄、病理类型及分化程度均无明显关系 (P >0 0 5 ) ;其血浆VEGF165浓度与肿瘤的TNM分期及远处转移有密切关系 (P <0 0 5 ) ,与淋巴结转移无关 (P >0 0 5 ) ;其外周血DC含量与肿瘤的TNM分期、淋巴结转移及远处转移均有密切关系 (P <0 0 5 )。体外实验显示 :与对照组相比 ,VEGF165实验组细胞中CD+ 14 细胞比例增高 ,DC表面CD40 、CD86、人类白细胞?

Objective To detect the effect of vascular endothelial growth factor (VEGF)on dendritic cells(DC)in patients with non-small cell lung cancer(NSCLC). Methods The measurement of DC in the peripheral blood was performed by a novel flow cytometric assay in 85 patients with NSCLC and 14 healthy volunteers. Enzyme-linked immunosorbent assay (ELISA) was used to measure the concentration of VEGF_ 165 in the plasma. CD +_ 14 peripheral blood mononuclear cells(PBMC) were cultured to obtain DC in vitro with cytokines. VEGF_ 165 was added to evaluate its effect on DC differentiation and survival. The phenotypes and apoptosis of cultured cells were detected by flow cytometry. Results In comparison with healthy volunteers, the level of VEGF_ 165 was significantly increased ( P< 0.05), while that of DC was significantly decreased ( P< 0.01) in patients with NSCLC. No significant correlation was noted between the concentration of VEGF_ 165 and age, gender, differentiation and histological types in patients with NSCLC, neither was found in the level of DC ( P> 0.05). The concentration of VEGF_ 165 was closely associated with TNM stage and distal metastasis ( P< 0.05), while no correlation was found between the concentration of VEGF_ 165 and lymph node metastasis ( P> 0.05). Significant correlations were noted between the level of DC in patients with NSCLC and TNM stage, lymph node metastasis and distal metastasis ( P< 0.05). There was a negative correlation between the concentration of VEGF_ 165 and the level of DC ( P< 0.05). Patients with abnormally elevated VEGF_ 165 showed significantly fewer DC. Cells cultured in vitro in the presence of VEGF_ 165 exhibited higher expression of CD +_ 14 ( P= 0.000) and increased ratio of apoptic cells ( P< 0.01), but decreased expression of CD_ 40 , CD_ 86 and HLA-DR( P< 0.01), as compared to cells cultured without VEGF_ 165 . Conclusions The level of DC and the concentration of VEGF in the peripheral blood can reflect the malignancy of NSCLC. NSCLC can over-express VEGF to inhibit DC differentiation and maturation to evade host immune surveillance.