HA纳米载体转hGM-CSF基因的HepG2疫苗的抗肿瘤活性研究

The Anti-Tumor Activity of HepG2 Tumor Vaccine Transfected hGM-CSF Gene Mediated by HA Nanoparticles

目的:评价HA纳米载体转hGM-CSF基因的HepG2疫苗的抗肿瘤活性。方法:SCID小鼠20只腹腔内注射健康志愿者外周血淋巴细胞(1×107/ml)1.0 ml,同时背部皮下接种人肝癌HepG2细胞(1×107ml)0.2 ml。当皮下移植瘤体积长至100 mm3时,随机分四组:Ⅰ组,60Co照射的转染GM-CSF基因的HepG2细胞组,Ⅱ组,60Co照射的HepG2细胞组,Ⅲ组,生理盐水组,Ⅳ组,接种前,每组5只。MTT法检测各组小鼠脾细胞CTL活性,ELISA法检测血清IL-4和IL-12的水平。结果:转染GM-CSF基因的HepG2疫苗组小鼠脾细胞CTL活性明显高于其余组(P<0.05);同时血清Th1类细胞因子IL-12的水平明显升高(P<0.05),而Th2类细胞因子IL-4的水平无统计学意义(P>0.05)。结论:HA纳米载体转hGM-CSF基因的HepG2疫苗可刺激机体产生特异性免疫反应。

Objective： To investigate the anti-tumor effects and mechanism of HepG2 tumor vaccine transfected hGM-CSF gene mediated by HA nanoparticles.Methods： Health human peripheral blood lymphocytes were isolated from peripheral blood of healthy donors and intraperitoneal injected into SCID mice at 2×10＆7/ ml.Simultaneously,hepatocellular carcinoma cells were intracutaneously injected into SCID mouse at 2×10^7/ ml.When the xenografted to a 100 mm3,mice was randomly divided into 4 groups： Group 1,HepG2 cells transfected with GM-CSF Gene after 60Co irradiation group;Group 2,HepG2 cells after 60Co irradiation group;Group 3,normal saline group： Group 4,uninjected group.Every group had 5 mice.Cytotoxic rates were detected by using MTT assay.The level of IL-12 and IL-4 in serum was detected by ELISA method.Results： The activaty of CTL were significantly higher in group HepG2 tumor vaccine transfected hGM-CSF gene mediated by HA nanoparticles than those in other groups（P〈0.05）;Serum level of Th1-type cytokine IL-12 increased after vaccination（P〈0.05）,whereas Th2-type cytokine IL-4 showed no significant changes（P〉0.05）.Conclusion： The HepG2 tumor vaccine transfected hGM-CSF gene mediated by HA nanoparticles can induce immune response.