HA纳米载体介导转hGM-CSF基因的HepG2疫苗诱导的抗瘤效应研究

Anti-tumor effects of HepG2 tumor vaccine transfected hGM-CSF gene mediated by HA nanoparticles

目的:观察HA纳米颗粒载体介导转染hGM-CSF基因的HepG2细胞疫苗体外抗肿瘤效应,为hGM-CSF基因修饰的HepG2细胞疫苗的临床应用提供依据。方法:HA纳米颗粒载体介导hGM-CSF基因转染HepG2细胞制备转GMCSF基因的HepG2细胞疫苗。密度梯度离心法分离人PBMC,体外诱导人PBMC。WST-1法测定PBMC的增殖活性及对HepG2细胞的杀伤效应,流式细胞术分析CD4+和CD8+的阳性表达率,ELISA法测定INF-γ的分泌。结果:WST-1结果显示,转基因HepG2组疫苗能诱导PBMC增殖,其增殖率优于野生型疫苗(p<0.05);其诱导的PBMC对HepG2的杀伤率高于各野生型疫苗组和各空白对照组(p<0.05)。FCM结果显示,转基因HepG2疫苗组PBMC中CD4+和CD8+阳性表达率均高于各野生型疫苗组和各空白对照组(p<0.05)。ELISA结果显示,转基因组PBMC培养上清中IFN-γ含量为1989.76±254.21pg/ml,高于各野生型疫苗组和各空白对照组(p<0.05)。结论:HA纳米颗粒载体介导转染hGM-CSF基因能增加HepG2细胞疫苗的免疫原性,转hGM-CSF基因HepG2细胞疫苗可有效诱导PBMC增殖、分化,增加INF-γ的分泌,提高其对HepG2细胞的杀伤作用。

Objective：To observe the HA Nanoparticles mediated transfection of hGM-CSF gene in HepG2 cells in vitro anti-tu-mor effect of the vaccine for the hGM-CSF gene-modified vaccine HepG2 cells provide the basis for clinical application.Methods：HA nanoparticles mediated gene transfer hGM-CSF preparation of HepG2 cells HepG2 cells transfected GMCSF gene vaccine.Density gra-dient centrifugation were PBMC,in vitro induction of human PBMC.WST-1 method PBMC proliferation activity and killing effect on HepG2 cells,flow cytometry analysis of CD4＋and CD8＋ positive expression rates,ELISA determination of INF-γ secretion.Results：WST-1 showed that transgenic HepG2 group of vaccine induced PBMC proliferation,the proliferation rate than wild-type vaccine（p0.05）.The PBMC of the HepG2 induced high anti-vaccine groups in the wild type and the control group（p0.05）.FCM results showed that transgenic HepG2 PBMC in the vaccine group CD4＋and CD8＋expression was higher than the wild-type vaccine group and con-trol group（p0.05）.ELISA results showed that the transgenic group PBMC supernatants of IFN-γ content was 1989.76±254.21 pg/ml,higher than the wild-type vaccine group and control group（p0.05）.Conclusion：HA nanoparticles mediated transfection of hGM-CSF gene can increase the immunogenicity of the vaccine HepG2 cells,transfected HepG2 cells in hGM-CSF gene vaccine can effectively induce PBMC proliferation,differentiation,increased INF-γ secretion,increase its HepG2 cells in vitro.