近年研究结果 显示,含有非甲基化的CpG基序具有强大的免疫激活作用[1].前期研究表明,CpG ODN可增强树突状细胞(DC)对胃癌细胞株MKN45的杀伤作用[2].本研究旨在观察CpGODN诱导DC的抗胃癌效应.

Objective To explore whether CpG ODN1826 assist dendritic cells(DC) to affect the anti-gastric cancer effects in vitro.Methods DC was isolated from normal human peripheral blood by GM-CSF+IL-4,and append TNF-α in the fifth and divided Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅴ,Ⅵ,Ⅶ,Ⅷ groups,and append carcinoma antigen 50 μl in every group,CpG ODN1826 10 μg/ml in Ⅱ,Ⅴ,Ⅵ,Ⅷ groups in the sixth day.IL-12 and IFN-γ levels were detected using ELISA in 10th day.The cytotoxicity of CTL to MKN45,MKN28,SGC7901,A549 was detected by MTT assay in vitro.The inhibitive effects of CpG ODN1826 on gastric cell proliferation cycle and apoptosis in vitro were detected by flow cytometry.Results On the 10th day,IL-12,IFN-γ level were significantly increased in the CpG ODN1826 groups.The CTL had much obviously raised the cytotoxicity to different differentiated types of the three gastric cancer cell lines such MKN45,MKN28,SGC7901 by CpG ODN1826,and than A549 (P<0.01).Treated with the CpG ODN1826 in vitro,the percentages of G0/G1,S and G2/M cells of tumor cells were(MKN45 68.35%,MKN28 69.23%,SGC790169.80%),(MKN45 39.45%,MKN28 39.75%,SGC7901 39.55%) and(MKN45 6.50%,MKN28 6.30%,SGC7901 6.42%) respectively.and the apoptosis of MKN45 75.20%,MKN23 73.87%,SGC7901 72.43%,A549 51.43%,indicated CpG ODN1826 may significantly inhibit the percentages of three gastric cell lines,and promote gastric cell early apoptosis(P<0.01).Conclusions The effecience and specificity of CpG ODN1826 can observably enhance dendritic cells induce the effecience and specificity of anti-gastric cancer activity,can significantly inhibit gastric cells proliferation and growth,and promote gastric cell early apoptosis,and might provide a new kind of therapeutic means for gastric cancer.

目的探讨树突状细胞（DC）抗胃癌效应的作用。方法分离正常人外周血单个核细胞（PBMC）,加入GMCSF和IL-4培养,于第5d加入TNF-α继续培养,诱导分离DC。将淋巴细胞与DC混合培养4d后,加入胃癌细胞MKN45、MKN28及SGC7901抗原培养至10d,诱导产生肿瘤特异性的CTL。ELISA检测IL-12和IFN-γ的水平,MTT法检测DC诱导的CTL对MKN45、MKN28、SGC7901和A549体外杀伤效应,流式细胞术检测CTL对肿瘤细胞周期、凋亡的影响。结果培养10d后,IL-12、IFN-γ的分泌量明显提高,DC对同种不同分化类型的胃癌细胞株MKN45、MNK28和SGC7901均有强烈的杀伤效应,杀伤活性显著高于A549（P＜0.01）。与A549比较,MKN45、MKN28和SGC7901的细胞周期显著抑制、凋亡率增加（P＜0.01）。结论 DC体外可诱导出高效而特异的抗胃癌效应,显著抑制胃癌细胞分裂增殖,促进其凋亡。