胃癌细胞-树突状细胞融合疫苗对肿瘤细胞增殖周期影响的研究

Inhibitive effects of gastric cancer cell-dendritic cell fusion vaccine on tumor cell proliferation cycle

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 study the inhibitive effects of gastric cancer cell-dendritic cell fusion vaccine on tumor cells of proliferation cycle. Methods Peripheral blood mononuclear cells were separated from gastric cancer patients and co-cultured with granulocyte macrophage colony stimulating factors （GM-CSF）, interleukin-4（IL-4） and tumor necrosis factur-α（TNF-α） to generate mature dendritic cells. The dendritic cells and SGC7901 cells were fused by polyethylene glycol, and the pure fusion cells were screened out by selective culture systems. The inhibitive effects of gastric cancer cell-dendritic cell fusion vaccines on tumor cell proliferation cycle in vivo and in vitro were detected by flow cytometry. Results Treated with the fusion vaccine in vitro, the percentages of G0/G1, S and G2/M cells of tumor cells were （76. 77 ±4. 38）%, （16. 50±2.90）% and （6. 73±1.59）% respectively. There were significant differences in the percentages of different cell cycle tumor cells between the tumor cells treated with the fusion vaccine and those co-cultured with dendritic cell or T cells alone（ P 〈 0.01 ） . The proliferative index of the tumor cells treated with the fusion vaccine was 23.34±3.51, significantly lower than those co-cultured with dendritic cell and controls （ P 〈 0.05）. Conclusions Fusion vaccines can affect cell cycle of the tumor cells, thus inhibit tumor cell proliferation and growth.