论文部分内容阅读
目的为胃癌细胞-树突状细胞(DC)融合瘤苗免疫治疗胃癌患者提供实验依据及理论基础。方法采用细胞融合技术制备胃癌细胞-DC融合疫苗,对融合疫苗影响体外培养肿瘤细胞增殖周期及体内种植瘤组织细胞周期的特点进行研究。(1)胃癌患者外周血单个核细胞经粒细胞-巨噬细胞刺激因子(GM-CSF)、白介素(IL)-4、肿瘤坏死因子(TNF)-α诱导分化获取DC；(2)DC与SGC7901细胞经聚乙二醇诱导融合,HAT/HT选择培养获得纯净融合细胞；(3)流式细胞术检测融合疫苗对体外培养胃癌细胞周期及体内种植瘤组织细胞周期的影响。结果(1)外周血单个核细胞诱导分化可获得具备典型特征及细胞表型的DC；(2)DC与SGC7901细胞经PEG诱导融合,HAT/HT选择培养获得纯净融合细胞；(3)体内应用融合疫苗组癌细胞周期比例：G_0/G_1间期(76．77±4．38)%,S期(16．50±2．90)%,G_2/M期(6．73±1．59)%；与对照组比较,P＜0．01,差异有统计学意义。体内应用融合疫苗组种植瘤组织细胞增殖指数为23．34±3．51,与对照组(65．73±4．43)比较,P＜0．05,差异有统计学意义。结论融合细胞疫苗对肿瘤细胞增殖周期可产生显著影响,明显减慢肿瘤生长速度,抑制肿瘤细胞分裂增殖,是融合细胞疫苗发挥更强生物学效应的基础之一。
Objective To provide experimental evidence and theoretical basis for the immunotherapy of gastric cancer patients with gastric cancer cell-dendritic cell (DC) fusion vaccine. Methods The gastric cancer cell-DC fusion vaccine was prepared by cell fusion technique. The effects of the fusion vaccine on the proliferation of tumor cells in vitro and the cell cycle of the implanted tumor in vivo were studied. (1) Peripheral blood mononuclear cells from gastric cancer patients were induced to differentiate by GM-CSF, IL-4 and TNF-α; (2) SGC7901 cells were induced by polyethylene glycol and pure fusion cells were obtained by HAT / HT selective culture. (3) Flow cytometry was used to detect the effect of fusion vaccine on the cell cycle of gastric cancer cells and the cell cycle of the implanted tumor in vivo. Results (1) DC with typical characteristics and cell phenotype can be obtained by differentiation of peripheral blood mononuclear cells; (2) DC fused with SGC7901 cells induced by PEG and pure fused cells were selected by HAT / HT selection; (3) The proportion of cancer cell cycle in fusion vaccine group was 76.77 ± 4.38% in G_0 / G_1 phase, 16.50 ± 2.90% in S phase, 6.73 ± 1.59 in G_2 / M phase, %; Compared with the control group, P <0.01, the difference was statistically significant. The proliferation index of the implanted tumor in the fusion vaccine group in vivo was 23.34 ± 3.51, which was significantly lower than that of the control group (65.73 ± 4.43), P <0.05. Conclusion The fusion cell vaccine can have a significant effect on the proliferation cycle of tumor cells, significantly slowing the growth of tumor cells and inhibiting the proliferation and proliferation of tumor cells. It is one of the foundations that the fusion cell vaccine exerts a stronger biological effect.