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为了探讨人白细胞介素 (h IL ) - 1 8基因转导的抗肿瘤作用 ,构建了 h IL- 1 8c DNA真核表达质粒载体 pc DNA3/h IL- 1 8.将 pc DNA3/h IL- 1 8经脂质体包裹 ,按照 1 0 0μg/只的剂量注射入雄性Balb/c小鼠后肢肌肉 ,在设定时间处死小鼠取注射部位肉提取总 RNA,应用半定量反转录聚合酶链反应 (RT- PCR)及免疫组化法检测了 h IL- 1 8m RNA及其蛋白质在小鼠肌肉中不同时间点的表达水平 .随后 30只雄性 Balb/c小鼠于基因注射后第 7d腹部皮下 (i.d.)或腹腔内 (i.p.)接种 1×1 0 5个同系小鼠 S- 1 80肉瘤细胞 .观察了 S- 1 80肿瘤细胞在腹腔及皮下的生长情况、小鼠体重、肿瘤重量及存活时间 .结果发现 ,pc DNA3/h IL- 1 8注射后 2 d能检测到 h IL- 1 8m RNA表达 ,第 7d表达量较高 ,第 2 8d仍有 h IL- 1 8m RNA表达 .免疫组化染色显示了注射部位散在有 h IL- 1 8蛋白质颗粒 .h IL- 1 8基因注射组小鼠体重、肿瘤重量均比对照组轻 (P值分别小于 0 .0 0 5、0 .0 5) ,存活时间比对照组延长 .结果显示 ,真核表达系统 pc DNA3/h IL - 1 8经脂质体包裹肌注后能有效表达 ,具有明显的抑制肿瘤生长作用 .
To investigate the antitumor effect of human interleukin (hIL) - 18 gene transduction, a hIL-8 8c DNA eukaryotic expression plasmid vector pc DNA3/h IL-1 was constructed. 8. pc DNA3/h IL- 1 8 was encapsulated by liposome and injected into the muscle of the hind limbs of male Balb/c mice at a dosage of 100 μg/body. The mice were sacrificed at the set time to take the total RNA extracted from the injection site and semi-quantitative reverse transcription polymerase was used. RT-PCR and immunohistochemistry were used to detect the expression level of hIL-8 mRNA and its protein in mouse muscle at different time points. The following 30 male Balb/c mice were injected at 7 days after gene injection. 1×10 5 syngeneic S-180 sarcoma cells were inoculated subcutaneously (id) or intraperitoneally (ip). Growth of S-180 tumor cells in the abdominal cavity and subcutaneous tissue, weight of the mice, and tumor weight were observed. And the survival time. The results found that pc DNA3/h IL-8 can be detected 2 h after injection of h IL-1 8m RNA expression, 7d high expression, the second 8d still h IL-8 8m RNA expression. Immunohistochemical staining showed that there were hIL-18 protein particles scattered at the injection site. The body weight and tumor weight of the mice injected with hIL-8 gene were compared. The group was lighter (P values were less than 0.05 and 0.5), and the survival time was longer than that of the control group. The results showed that the eukaryotic expression system pc DNA3/h IL-18 could be intramuscularly injected with liposomes. Effective expression, has a significant inhibitory effect on tumor growth.