中文摘要：

目的 制备肿瘤相关糖蛋白-72（TAG-72）嵌合锚定T细胞,体内外实验检测其对肝癌的抑癌活性.方法 分离健康人外周血单个核细胞（PBMC）,采用脂质体lipofectamineTM 2000介导的细胞转染,将已制备的重组真核表达载体抗TAG-72 scFv-CD3ξ-pcDNA3.0导入PBMC,获得肿瘤相关糖蛋白-72（TAG-72）嵌合锚定T细胞,以此为实验组治疗细胞;用SDS-PAGE检测TAG-72嵌合锚定T细胞中抗TAG-72 scFv-CD3ξ嵌合分子的表达,并用Western blotting证实.用转染空载体pcDNA3.0的PBMC作为对照组治疗细胞.将实验组及对照组的治疗细胞分别经尾静脉注射治疗荷TAG-72＋肝癌细胞HepG2、荷TAG-72肝癌细胞BEL7402裸鼠动物模型,观察对裸鼠的抑癌效应.结果 SDS-PAGE检测TAG-72嵌合锚定T细胞嵌合分子抗TAG-72 seFv-CD3ξ的表达,显示大小为47 kDa的融合蛋白片段,与抗TAG-72 scFv-CD3ξ的理论值相符;Western blotting证实该片段CD3呈阳性表达.TAG-72嵌合锚定T细胞与肝癌细胞HepG2共培养后可对后者产生G1期阻滞.用TAG-72嵌合锚定T细胞经尾静脉注射治疗荷瘤裸鼠,发现实验组细胞对荷HepG2细胞裸鼠存活时间、体质量、肿瘤包块大小等方面疗效优于荷BEL7402细胞裸鼠的疗效,两者比较差异有统计学意义（P＜0.05）.结论 TAG-72嵌合锚定T细胞可特异性抑制荷TAG-72＋肝癌细胞裸鼠的肝癌细胞生长;该治疗策略将为肝癌的临床诊治提供一种有希望的途径.

英文摘要：

Objective To prepare tumor associated glycoprotein-72 （TAG-72） targeted T lymphocytes,and to study the antitumor effect of it for hepatocellular cancer.Methods Peripheral blood mononuclear cells （PBMC） were isolated from healthy volunteers,the recombinant vector anti-TAG-72-scFv-CD3ξ-pcDNA3.0 were transfected into PBMCs by lipofectamine TM 2000,TAG-72 special lymphocytes were obtained.Meanwhile,lymphocytes transfected with plasmid pcDNA3.0 were used as controls.TAG-72 special lymphocytes and that of controls were injected by tail vein of hepatocellular cancer-bearing nude mice.Antitumor response was observed.Results T lymphocytes were cultivated 24 hours and the expression of anti-TAG-72 scFv-CD3ξ was detected by SDS-PAGE and Western blotting.A 47 kDa protein was confirmed as theoretical analysis and proved CD3 ＋ T lymphocytes were cultivated after 12 and 24 hours,hepatocellular carcinoma cells were co-cultured with them.G1 blocks of TAG-72 ＋ hepatocellular tumor HepG2 cells were observed.Annexin V-FITC/PI double staining FCM assay was used to further determine the TAG-72 ＋/ hepatocellular tumor cocultivated with T lymphocytes with chimeric receptor.The apoptotic rate of TAG-72 ＋ hepatocellular tumor HepG2 cells was （47.8 ±4.2）％,while that of TAG-72 hepatocellular tumor BEL7402 cells was （18.7 ±2.8）％,which was significantly lower than that of TAG-72 ＋ hepatocellular tumor cells.Conclusion This receptor design will facilitate efficient generation of genetically T lymphocytes with chimeric receptor and may provide valuable tools for the cellular immunotherapy of hepatocellular cancer.