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目的研究胃癌细胞培养上清液对树突状细胞(DCs)表型的影响。方法酶联免疫法检测胃癌及正常胃上皮细胞上清液中血管内皮生长因子(VEGF)、白细胞介素(IL)-10和肿瘤生长因子(TGF)-β1蛋白含量。自健康人外周血获取DCs,流式细胞仪(FCM)检测加入上述细胞培养上清液后DCs表型CD83、CD80、CD86、HLA-DR的变化。结果OCUM-2MD3、BGC-823和HGC-27细胞株的胃癌细胞培养上清液中均可检测到VEGF蛋白;IL-10、TGFβ-1蛋白在OCUM-2MD3、BGC-823细胞株中有表达,在HGC-27中未见表达。镜下观察细胞具有DCs典型的形态特征,单纯用GM-CSF和IL-4培养的细胞低表达HLA-DR、CD80、CD86和CD83,加入肿瘤坏死因子(TNF)α-后,表达水平明显上调。加入细胞培养上清液后,FCM检测不同胃癌细胞组各表型表达均显著降低,而正常胃上皮细胞组无明显变化。结论应用GM-CSF、IL-4和TNF-α体外刺激,可成功扩增出成熟DCs。胃癌细胞可通过分泌VEGF、IL-10、TGF-β1等抑制DCs成熟,降低其抗原提呈功能,而进行免疫逃逸。
Objective To investigate the effect of gastric cancer cell culture supernatant on the dendritic cells (DCs) phenotype. Methods The levels of vascular endothelial growth factor (VEGF), interleukin (IL) -10 and TGF-β1 in gastric cancer and normal gastric epithelial cell supernatant were detected by enzyme-linked immunosorbent assay. DCs were obtained from peripheral blood of healthy volunteers, and the changes of CD83, CD80, CD86 and HLA-DR of DCs were detected by flow cytometry (FCM) after adding the above cell culture supernatant. Results The VEGF protein was detected in the culture supernatants of gastric cancer cells of OCUM-2MD3, BGC-823 and HGC-27 cell lines. The expressions of IL-10 and TGFβ-1 proteins were found in OCUM-2MD3 and BGC-823 cell lines , No expression in HGC-27. Microscopically, the cells showed the typical morphological features of DCs. The cells cultured with GM-CSF and IL-4 alone showed low expression of HLA-DR, CD80, CD86 and CD83. The expression of TNF-α was significantly up-regulated . After addition of cell culture supernatant, the expression of FCM in different gastric cancer cell groups was significantly decreased, while the normal gastric epithelial cell group had no significant change. Conclusion Mature DCs can be successfully amplified by GM-CSF, IL-4 and TNF-α in vitro. Gastric cancer cells can escape immune evasion by inhibiting the maturation of DCs by secreting VEGF, IL-10 and TGF-β1 and reducing their antigen presenting function.