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目的观察共表达基因疫苗pc IL-18/MAGE-1对绒癌JEG-3细胞株HLA-G基因表达的影响,探究pc IL-18/MAGE-1抗肿瘤作用机制。方法应用构建的共表达pc IL-18/MAGE-1 DNA疫苗免疫小鼠,同时设阴性对照组和空白对照组,免疫后分别收集脾细胞,作用于靶细胞JEG-3,应用FCM检测小鼠T细胞亚群情况及NK细胞活性,MTT法检测肿瘤特异性CTL对靶细胞的杀伤率,RT-PCR法检测HLA-G基因表达情况。结果免疫组HLA-G基因表达降低,且对JEG-3细胞杀伤活性增加,与对照组比较,差异显著(P<0.05)。免疫组NK细胞活性及CD4+/CD8+、CD8+、CD4+值均高于对照组(P<0.05)。结论共表达基因疫苗pc IL-18/MAGE-1通过降低HLA-G基因的表达,进而增加NK细胞活性以及诱导肿瘤特异性CTL直接杀伤肿瘤细胞,发挥抗肿瘤作用。
Objective To investigate the effect of pc IL-18 / MAGE-1 co-expressing gene vaccine on the expression of HLA-G gene in choriocarcinoma JEG-3 cells and explore the antitumor mechanism of pc IL-18 / MAGE-1. Methods The recombinant plasmid pc IL-18 / MAGE-1 was used to immunize mice. At the same time, negative control group and blank control group were immunized with spleen cells respectively. The spleen cells were collected and treated with JEG-3. FCM T cell subsets and NK cell activity. The killing rate of tumor-specific CTL on target cells was detected by MTT assay. The expression of HLA-G gene was detected by RT-PCR. Results The expression of HLA-G gene in the immunized group was decreased and the cytotoxicity against JEG-3 cells was increased. Compared with the control group, the difference was significant (P <0.05). The NK cell activity and CD4 + / CD8 +, CD8 + and CD4 + values in the immunized group were higher than those in the control group (P <0.05). Conclusions The co-expression of gene vaccine pc IL-18 / MAGE-1 exerts anti-tumor effect by decreasing the expression of HLA-G gene, further increasing the activity of NK cells and inducing tumor-specific CTL to directly kill tumor cells.