MAGE-1修饰的树突状细胞体外诱导杀伤人肝癌细胞  

Study of the cytotoxity against human hepatocellular carcinoma cells induced by the MAGE-1 gene modified dendritic cells

作　　者：刘彬彬[1] 叶胜龙[1] 贺平[1] 郑宁[1] 赵燕[1] 孙瑞霞[1] 汤钊猷[1] 

机构地区：[1]上海医科大学中山医院肝癌研究所,200032

出　　处：《中华肝脏病杂志》2001年第3期151-153,共3页Chinese Journal of Hepatology

基　　金：国家自然科学基金(39770826);美国中华医学基金会(CMB)部分资助(93583)

摘　　要：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 genetically modify dendritic cells (DC) with a tumor associated antigen gene MAGE-1 and to observe in vitro the cytotoxic effect induced by this genetically modified DC against human hepatocellular carcinoma (HCC) cell line SMMC7721, thus giving a primary verification of the possibility of this genetically modified DC to induce specific antitumor immunity against HCC and serving as a new type of vaccine. Methods The MAGE-1 gene was inserted into the retrovirus vector LXSN to construct the recombinant retrovirus LMSN. The monocyte-derived DCs were transfected by LMSN and control virus LXSN respectively as well as a third group was set up as non-transfected control. The MAGE- 1 gene expression in LMSN transfected DC was identified by Western blot and the in vitro cytotoxities against SMMC7721 induced by three groups of DC were tested by MTT assay. Results A recombinant retrovirus LMSN containing the tumor rejection antigen gene MAGE- 1 was successfully constructed and the MAGE-1 gene expression in LMSN transfected human monocyte-derived DC was induced. The LMSN transfected DC could induce very high lytic activity against SMMC7721 in vitro with the lytic activity of 78.9% ± 3.6%. Nevertheless, the LXSN transfected DC and non-transfected DC only induced relatively lower lytic activity as 34.7% ± 4.3% and 3.9%±2.0%, respectively. The difference of the lytic activities between those three groups as statistically significant (P<0.01). Conclusions The MAGE-1 gene transfected DC can induce higher in vitro cytotoxicity against SMMC7721, suggesting the possibility of this genetically modified DC to induce specific antitumor activity and to serve as a new type of vaccine for HCC.

关 键 词：树突细胞 基因治疗 肝细胞癌 MAGE-1 体外诱导 

分 类 号：R735.7[医药卫生—肿瘤]