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.

Objective To explore whether CpG ODN1826 can affect the antitumor activity of dendritic cells ( DC) against gastric cancer in vitro. Methods DC was isolated from normal human peripheral blood by GM-CSF + IL-4,GM-CSF + IL-4 + TNF-α,GM-CSF + IL-4 + LPS and GM-CSF + IL-4 + CpG ODN. The surface signal of DC was analysed by fluorescence-actived cell sorting( FACS) ,and the abilities to stimulate proliferation of allogenic lymphocyte by DC and antitumor experiment were detected by MTT assay,and IL-12 released by every DC was detected by ELISA. Results A number of cells had typical morphology characteristics on the 10th day of CpG ODN group and LPS group,the CpG ODN group cells expressed the highest level of CD40,CDla,CD80,CD86,MHC-Ⅱand secretion of IL-12,and was potent to stimulate the proliferation of allogenic lymphocytes,and enhanced the antitumor activity of DC. Conclusions CpG ODN may evidently enhance the differentiation and maturation of DC and increase the antitumor capacity by stimulating PBMC in vitro.