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目的 探讨白细胞介素 (IL) 12基因转染人胰腺癌细胞对肿瘤的抑制作用及机制。方法 用逆转录病毒MFG作为载体 ,通过逆转录PCR和磷酸钙沉淀法将鼠IL 12基因转染导入人胰腺癌细胞PK 1,得到PK1/IL 12细胞。单纯用MFG感染PK1所得细胞命名为PK1/MFG。用ELISA方法检测IL 12产生量 ,用NK细胞抗体阻断NK细胞作用 ,比较活体及离体条件下两种细胞的增长曲线。活体显微镜下观察肿瘤细胞的血管生成及IL 12的抑制效应。结果 PK1/IL 12细胞株的IL 12产生量为 1 12 μg·ml-1·48h-1。离体条件下两种细胞的生长曲线相似 ,但在活体条件下PK1/IL 12细胞的生长被显著抑制 (P <0 0 1) ,其血管生成被完全阻断 ,正常毛细血管亦被破坏。结论 IL 12基因转染可以抑制人胰腺癌细胞的生长。在免疫缺陷条件下 ,IL 12的抗血管生成作用足以抑制肿瘤细胞的血管生成及肿瘤的生长。IL 12的血管生成抑制效应缺乏特异性。
Objective To investigate the inhibitory effect and mechanism of human pancreatic cancer cells transfected with interleukin (IL) 12 gene. METHODS: Retroviral PCR and calcium phosphate precipitation were used to transfect murine IL 12 into human pancreatic cancer cells PK1 using retrovirus MFG as a vector to obtain PK1/IL 12 cells. The cells obtained by infecting PK1 with MFG alone were designated PK1/MFG. ELISA was used to detect the amount of IL-12 produced, and NK cells were used to block the effects of NK cells. The growth curves of the two cells were compared between live and ex vivo conditions. The angiogenesis of tumor cells and the inhibitory effect of IL 12 were observed under a living microscope. Results The production of IL 12 in PK1/IL 12 cell lines was 1 12 μg·ml -1·48h -1 . The growth curves of the two cells were similar under ex vivo conditions, but the growth of PK1/IL 12 cells was significantly inhibited in vivo (P < 0.01), angiogenesis was completely blocked, and normal capillaries were also destroyed. Conclusion IL 12 gene transfection can inhibit the growth of human pancreatic cancer cells. In immunodeficient conditions, the antiangiogenic effect of IL 12 is sufficient to inhibit tumor cell angiogenesis and tumor growth. The angiogenesis inhibitory effect of IL 12 lacks specificity.