论文部分内容阅读
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 investigate the effect and mechanism of cell culture supernatant on HepG2 cell apoptosis after transfection of human monocyte-derived dendritic cells derived from human hepatitis B core antigen recombinant plasmid pEGFP-N1 / CpG-HBcAg (ISS). Methods: The eukaryotic expression plasmid pEGFP-N1 / CpG-HBcAg (ISSa, c) was constructed and transfected into DCs from human peripheral blood. The apoptosis of HepG2 cells was induced by culture supernatants. The expression of CD80 and CD86 on the transfected DCs was detected by flow cytometry, and the apoptosis of HepG2 cells induced by culture supernatants was detected. The levels of IFN-γ, IL-2, IL-12, IL-4 and IL-10 in DC culture supernatants were detected by ELISA. Results: The expression of CD80 and CD86 in DCs transfected with pEGFP-N1 / CpG-HBcAg (ISSa) were significantly increased (P <0.01). After transfection, the expression of Th1 cytokines IFN-γ, IL-2 and IL-12 in supernatants were increased (P <0.01), and the expressions of Th2 cytokines IL-4 and IL-10 were decreased The culture supernatant of pEGFP-N1 / CpG-HBcAg (ISSa) group had a pro-apoptotic effect on HepG2 cells. With the prolongation of culture time, the apoptosis rate of HepG2 cells increased gradually. The apoptosis rate of HepG2 cells reached the maximum at 24 hours after induction 18.4%. Conclusion: The transfection of recombinant plasmid pEGFP-N1 / CpG-HBcAg (ISSa) can obviously promote the apoptosis of HepG2 cell line.