| 曾治中1 段晓明2△ 程元星1 黄璐1 贺修胜3.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[J].,2011,11(4):672-676 |
| 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 genemediated by HA nanoparticles |
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| DOI： |
| 中文关键词: 肝肿瘤 肿瘤疫苗 免疫治疗 粒细胞巨噬细胞集落刺激因子 外周血单个核细胞 |
| 英文关键词: liver neoplasms tumor vaccine immune therapy granulocyte-macrophage colony-stimulating factor peripheral blood
mononuclear cells |
| 基金项目:湖南省卫生厅科研基金资助项目(B2005-178) |
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| 中文摘要: |
| 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±254.21 pg/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。 |
| 英文摘要: |
| Objective: To observe the HA Nanoparticles mediated transfection of hGM-CSF gene in HepG2 cells in vitro anti-tumor
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 gradient
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 (p <0.
05). The PBMC of the HepG2 induced high anti-vaccine groups in the wild type and the control group (p <0.05). FCM results showed
that transgenic HepG2 PBMC in the vaccine group CD4 + and CD8 + expression was higher than the wild-type vaccine group and control
group (p <0.05). 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 (p <0.05). 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. |
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