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目的 通过观察肿瘤相关抗原基因 ＭＡＧＥ－１转导的树突状细胞（ｄｅｎｄｒｉｔｉｃ ｃｅｌｌｓ， ＤＣ）体外诱导对人肝癌细胞株ＳＭＭＣ７７２１的细胞毒作用，探讨基因工程ＤＣ诱导特异性抗肝癌免疫的能力及其作为新型肝癌瘤苗的可能性。方法用基因工程手段构建含ＭＡＧＥ－１基因的重组逆转录病毒。经包装后转染从人外周血单个核细胞（ＰＢＭＮＣ）诱导培养的ＤＣ， Ｗｅｓｔｅｒｎ ｂｌｏｔ方法鉴定ＭＡＧＥ－１基因的表达。用ＭＴＴ法检测转染及对照组ＤＣ体外诱导的对靶细胞的细胞毒作用，并计算杀伤率。结果 构建了一种含ＭＡＧＥ－１基因的重组逆转录病毒ＬＭＳＮ并转染了从人ＰＢＭＮＣ中诱导培养的ＤＣ。 Ｗｅｓｔｅｒｎ ｂｌｏｔ方法检测到外源ＭＡＧＫ－１基因在转染ＤＣ中的表达。 ＬＭＳＮ转染ＤＣ体外诱导的淋巴细胞在效靶比１０：１的情况下对靶细胞的杀伤率为７８．９％± ３．６％，ＬＸＳＮ转染ＤＣ诱导的杀伤率为３４． ７％±４．３％，而未转染ＤＣ诱导的杀伤率为３．９％±２１％。三组间的差别有显著性意义（Ｐ＜０．０１）。结论肿瘤相关抗原基因ＭＡＧＥ－１修饰的ＤＣ可在体外诱导出较强的对人肝癌细胞株ＳＭＭＣ７７２１的细胞毒作用。提示这种基因工程ＤＣ可能具诱导特?
Objective To investigate the cytotoxicity of human tumor cell line SMMC7721 induced by dendritic cells (DC) transduced with the tumor-associated antigen gene MAGE-1, and investigate the ability of genetically engineered DCs to induce specific anti-hepatocellular carcinoma immunity. The possibility of a new type of liver cancer vaccine. Methods The recombinant retrovirus containing MAGE-1 gene was constructed by genetic engineering. After packaging, DCs induced from human peripheral blood mononuclear cells (PBMNC) were transfected and Western blot was used to identify the expression of MAGE-1 gene. MTT assay was used to detect the cytotoxicity of target cells induced by transfection and control DCs in vitro, and the killing rate was calculated. RESULTS: A recombinant retrovirus LMSN containing the MAGE-1 gene was constructed and transfected with DCs induced from human PBMNCs. Western blot analysis detected the expression of exogenous MAGK-1 gene in transfected DCs. The killing rate of lymphocytes induced by LMSN-transfected DCs in vitro was 78.9%±3.6% when the ratio of target to target was 10:1. The killing rate of LXSN-transfected DCs was 34. 7% ± 4.3%, whereas untransfected DC induced a kill rate of 3.9% ± 21%. The difference between the three groups was significant (P<0.01). Conclusion The tumor-associated antigen gene MAGE-1 modified DC can induce strong cytotoxicity against human hepatoma cell line SMMC7721 in vitro. This suggests that this gene engineering DC may have induced special?