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目的:探讨核酸疫苗pVVP3IL-18HN对Hep-2肿瘤细胞的杀伤机制和对荷瘤动物模型肿瘤的抑制效应。方法:采用脂质体介导法将核酸疫苗pVVP3IL-18HN体外转染Hep-2细胞,运用吖啶橙/溴化乙锭(AO/EB)染色从细胞形态学角度观察肿瘤细胞的死亡方式,并进一步运用流式细胞仪(FCM)检测肿瘤细胞线粒体跨膜电位、活性氧水平和主要组织相容性复合体I类分子(MHC-I)表达情况。在C57BL/6小鼠右后肢皮下接种H22肿瘤细胞2×105个,荷瘤第7、14和21天,瘤内注射pVVP3IL-18HN,同时设pVVP3、pVIL-18、PVHN和空白对照组,计算抑瘤率,并检测细胞毒T淋巴细胞活性。结果:pVVP3IL-18HN转染可使Hep-2肿瘤细胞皱缩被AO/EB浓染,并上调肿瘤细胞活性氧水平和MHC-I分子表达,下调线粒体跨膜电位;对荷瘤小鼠模型肿瘤的抑瘤率为60.5%,与对照组比较,差异有统计学意义(P<0.05)。结论:pVVP3IL-18HN主要通过诱导细胞凋亡杀伤喉癌细胞,并对实体肿瘤有显著的抑制作用。
Objective: To investigate the mechanism of killing effect of DNA vaccine pVVP3IL-18HN on Hep-2 tumor cells and its inhibitory effect on tumor-bearing animal models. Methods: The recombinant plasmid pVVP3IL-18HN was transfected into Hep-2 cells by liposome-mediated method. The apoptosis of tumor cells was observed by acridine orange / ethidium bromide staining (AO / EB) Flow cytometry (FCM) was used to detect the expression of mitochondrial transmembrane potential, reactive oxygen species and major histocompatibility complex class I molecules (MHC-I) in tumor cells. In the right hindlimb of C57BL / 6 mice, 2 × 105 H22 tumor cells were inoculated subcutaneously, and pVVP3IL-18HN cells were injected intratumorally on the 7th, 14th and 21th day after tumor implantation, and pVVP3, pVIL-18, PVHN and blank control groups Inhibition rate, and detect cytotoxic T lymphocyte activity. Results: Transfection of pVVP3IL-18HN could induce the shrinkage of Hep-2 tumor cells by AO / EB, up-regulate the levels of reactive oxygen species (ROS) and MHC-I expression in tumor cells and down-regulate the transmembrane potential of tumor cells The inhibition rate was 60.5%, compared with the control group, the difference was statistically significant (P <0.05). Conclusion: pVVP3IL-18HN can kill laryngeal cancer cells mainly by inducing apoptosis, and has a significant inhibitory effect on solid tumors.