原发性肝癌基因治疗目的基因筛选的初步研究

The evaluation of objective genes for experimental gene therapy in primary hepatocellular carcinoma

目的 通过动物实验初步筛选对肝癌有明确疗效的目的基因。方法 应用携带对肿瘤有明确疗效的目的基因白细胞介素 2 (interleukin 2 ,IL 2 )、白细胞介素 12 (interleukin 12 ,IL 12 )、共刺激因子B7 1(costimulatorB7 1,B7 1)、单纯疱疹病毒胸苷激酶 (herpessimplex 1thymidinekinase ,HSV TK)、抑癌基因p5 3及白细胞介素 1β转换酶 (interleukin 1βconvertingenzyme,ICE)的逆转录病毒包装细胞系PA317,将该包装细胞系与肝癌细胞CBRH3 混合接种于大鼠腹腔中。结果 生理盐水及病毒载体对照组大鼠的平均生存时间分别为 (8 2± 0 8)及 (8 3± 1 1)d ,IL 2、B7 1、HSV TK、p5 3及ICE治疗组大鼠的平均生存时间分别为 (15 8± 2 4)、(10 6± 1 8)、(15 5± 2 5 )、(15 1± 2 5 )、(16 3± 2 0 )d ,生存期明显延长。IL 12治疗组的大鼠均存活超过 2个月。携带IL 12的逆转录病毒包装细胞株与CBRH3 的比例超过 1∶10时可使大鼠长期生存 (2个月观察期 ) ,而低于 1∶10 0时大鼠平均生存时间为(14 6± 2 1)d。结论 应用IL 12目的基因对肝癌的疗效最为显著。携带IL 12的逆转录病毒包装细胞株通过激活机体免疫细胞可完全杀灭超过其 10倍的肝癌细胞。

Objective[WT5”BZ] To select effective target gene to treat experimental hepatocellular carcinoma. [WT5”HZ]Methods[WT5”BZ] Retroviral packaging cell lines PA317 carrying the objective genes, including IL 2, IL 12, B 7 1, HSV TK, p53 and ICE,respectively were produced. PA317 and liver cancer cell line CBRH 3 were mixed in ratio of 1∶1 and inoculated into the peritoneal cavity of rats. The survival time of rats and the size of tumor were observed.[WT5”HZ] Results[WT5”BZ] The average survival time of rats in groups of normal saline and empty retroviral vectors were (8 2±0 8) and (8 3±1 1) days. The survival times in groups treated with IL 2, B 7, HSV TK, p53 and ICE were (15 8±2 4)、(10 6±1 8)、(15 5±2 5)、(15 1±2 5)、(16 3±2 0) days, which were much longer than in control groups.Rats treated with IL 12 all survived longer than two months when PA317 and CBRH 3 were mixed in a ratio larger than 1∶10.Decreasing the ratio to 1∶100 could shorten the survival time to (14 6±2 1) days.[WT5”HZ]Conclusions[WT5”BZ] Among the six genes used in liver cancer gene therapy, IL 12 was the most potent target gene. Followed up experiments proved that PA317 carrying IL 12 killed ten times as much cells of hepatocellular carcinoma through activating immunocytes in vivo.