重组质粒pEGFP-N1/CpG-HBcAg(ISS)转染DC培养上清诱导HepG2株凋亡的研究  

The study on apoptosis of liver cancer cell line HepG2 induced by culture supernatant of DCs transfection with recombinant plasmid pEGFP-N1/CpG-HBcAg(ISS)

作　　者：周健[1] 田德英[2] 许东[2] 张振纲[2] 陈淼 章述军[2] 吴会玲[2] 

机构地区：[1]武汉科技大学附属天佑医院感染科,武汉430064 [2]华中科技大学同济医学院附属同济医院感染科,武汉430030

出　　处：《中国免疫学杂志》2010年第6期488-493,共6页Chinese Journal of Immunology

基　　金：湖北省自然科学基金资助项目(No.2006ABA139)

摘　　要：目的:探讨人类乙肝核心抗原重组质粒pEGFP-N1/CpG-HBcAg(ISS)转染人外周血单核细胞来源树突状细胞后,细胞培养上清诱导肝癌细胞株HepG2凋亡的作用及机制。方法:构建真核表达质粒pEGFP-N1/CpG-HBcAg(ISSa,c),将其转染人外周血来源DC,用培养上清诱导HepG2的凋亡。用流式细胞仪检测已转染DC表面CD80和CD86的表达,检测培养上清诱导HepG2凋亡的变化。用ELISA法检测转染后DC培养上清的IFN-γ、IL-2、IL-12、IL-4和IL-10的水平。结果:pEGFP-N1/CpG-HBcAg(ISSa)转染DC表面CD80和CD86的表达均有明显升高(P<0.01)。转染后上清中Th1型细胞因子IFN-γ、IL-2和IL-12的表达增强(P<0.01),Th2型细胞因子IL-4和IL-10的表达下降(P<0.05),pEGFP-N1/CpG-HBcAg(ISSa)组培养上清对HepG2细胞具有促凋亡作用,随着培养时间延长,细胞凋亡率逐渐增加,HepG2细胞在诱导后24小时凋亡率达到最大,为18.4%。结论:重组质粒pEGFP-N1/CpG-HBcAg(ISSa)转染培养上清能明显促进肝癌细胞株HepG2的凋亡。Objective：To explore the effect of DCs transfected with recombinant plasmid pEGFP-N1/CpG-HBcAg（ISS） on inducing apoptosis of HepG2 cells and its mechanisms.Methods：Recombinant plasmid pEGFP-N1/ CpG-HBcAg（ISSa,c） was constructed and used to transfect DCs.The expression of CD80 and CD86 on the transfected DCs and the alteration of apoptosis HepG2 were analyzed by flow cytometry.IFN-γ,IL-2,IL-12,IL-4 and IL-10 in the culture supematant of the transfected DCs were detected by ABC-ELISA.Results：pEGFP-N1/CpG-HBcAg （ISSa）-transfected DCs expressed higher level of CD80 and CD86（P0.01）.The result showed that high level of Th1 type cytokines of IFN-γ,IL-2 and IL-12 were induced in pEGFP-N1/CpG-HBcAg（ISSa） recombinant plasmid groups（P0.01）,whereas the level of Th2 type cytokines such as IL-4 and IL-10 was inhibited obviously（P0.05）.The culture supernatant of pEGFP-N1/CpG-HBcAg （ISSa） group increased apoptosis rate of HepG2 with training time,and the highest apoptosis rate was 18.4% in the 24 h.Conclusion：The culture supernatant of pEGFP-N1/CpG-HBcAg （ISSa） group could significant increase apoptosis of HepG2.

关 键 词：CPG基序 树突状细胞 HEPG2 共刺激分子 TH1/TH2型细胞因子 

分 类 号：R323.47[医药卫生—人体解剖和组织胚胎学]