转铁蛋白-多聚乙烯亚胺介导IL-12基因对小鼠抗骨肉瘤免疫的增强作用

Enhancement of transferrin-polyethylenimine in mediating interleukin-12 gene on anti-osteosarcoma immunity in mice

目的:研究以转铁蛋白-多聚乙烯亚胺(transferrin-polyethylenimine,Tf-PEI)为靶向性载体,体内转染小鼠白细胞介素12(murineinter-leukin-12,mIL-12)基因治疗小鼠骨肉瘤模型的疗效。方法:分别以PEI和Tf-PEI为载体,将mIL-12基因导入体外培养的小鼠骨肉瘤细胞,并观察游离转铁蛋白对Tf-PEI转染效率的影响。建立小鼠骨肉瘤动物模型,在荷瘤部位将Tf-PEI包裹mIL-12基因直接注入肿瘤中,检测此基因在肿瘤细胞中的蛋白表达情况和小鼠脾脏自然杀伤细胞(NK)、细胞毒T淋巴细胞(CTL)的活性。结果:Tf-PEI组mIL-12蛋白表达效率显著高于PEI组(t=38.15,P<0.001)。游离转铁蛋白可显著抑制Tf-PEI组的转染效率(t=44.36,P<0.001)。在注射有mIL-12基因的肿瘤组织中,小鼠IL-12蛋白水平明显升高,为(72±5)ng/L(F=3.449,P<0.001),小鼠脾细胞NK,CTL活性增强(t=5.04,t=6.13,P<0.001)。结论:Tf-PEI是一种高效率的肿瘤靶向性基因转染载体,它可以成功的将mIL-12基因导入小鼠骨肉瘤模型,mIL-12基因治疗可提高机体的抗肿瘤免疫应答。

AIM:To investigate the effect of transferrin polyethylenimine(Tf PEI) as a tumor targeted carrier transfected in vivo murine interleukin 12(mIL 12) gene on the treatment of osteosarcoma model of mice. METHODS:PEI and Tf PEI were used as vectors.The mIL 12 gene was imported to the osteosarcoma cells of mice cultured in vitro,and the effect of free transferrin on the transfection efficiency of Tf PEI was observed.The animal model of osteoscarcoma in mice was established,Tf PEI mIL12 plasmid complexes were injected to the tumor directly,and the expressional condition of protein of this gene in the tumor cells,the activity of natural killer(NK) cells,and cytotoxic T lymphocyte(CTL) of spleen in mice were detected. RESULTS:The expression efficiency of mIL12 in Tf PEI group was significantly higher than that in PEI group(t =38.15,P< 0.001),the free transferring could inhabit the transfection efficiency of Tf PEI obviously(t=44.36,P< 0.001). The level of IL 12 in mice was increased obviously(72±5) ng/L(F=3.449,P< 0.001) in the tumor tissue which was injected with mIL12.The activities of NK and CTL were increased in spleen of mice(t=5.04, t=6.13, P< 0.001). CONCLUSION:Tf PEI is a high efficiency tumor targeted vector.It can successfully import mIL 12 gene into the mice model of osteosarcoma,and mIL 12 gene therapy is able to promote the immune response of anti tumor efficiently.