摘要
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 research the antitumor efficacy on mice model bearing H22 tumor with application of pVVP3IL-18HN and its mechanism of the antitumor effect on Hep-2.Method:pVVP3IL-18HN was introduced into Hep-2 cells by liposome,then cellular morphology was observed through AO/EB stain.To evaluate the mechanism of the antitumor effect on Hep-2 of pVVP3IL-18HN and its effect on tumor immunogenicity, flow cytometer(FCM) was used to detect the variation of mitochondrial trans-membrane potential,reactive oxygen species(ROS) and the MHC-I.C57BL/6 mice were subcutaneously inoculated with 2×10~5 H22 tumor cells in the right hindlimb,and 7,14 and 21 day post-inoculation,pVVP3IL-18HN was administrated intratumorally.The pVVP3,pVIL-18,PVHN and PBS control group were treated as above.Then the tumor suppression and specific cytotoxic T lymphocyte(CTL) activity was detected.Result:After the transfection of pVVP3IL-18HN,tumor cells crenated and stained by AO/EB. In the research pVVP3IL-18HN was found to up-regulate the value of ROS and the expression of MHC-I,and down-regulate the value of mitochondrial trans-membrane potential. The tumor suppression rate of pVVP3IL-18HN was 60.5% and a dramatically increased tumor suppression was revealed in the pVVP3IL-18HN treated mice as compared with control mice treated with PBS(P<0.05).Conclusion:pVVP3IL-18HN kill the tumor cells by apoptosis and have notable antitumor effect in vivo.
出处
《临床耳鼻咽喉科杂志》
CAS
CSCD
北大核心
2005年第15期704-706,共3页
Journal of Clinical Otorhinolaryngology
基金
"973"规划项目(No:G199011902)
自然科学基金重大项目(No:39893290-4-3)
关键词
核酸疫苗
喉肿瘤
细胞凋亡
pVVP3IL-18HN
Laryngeal neoplasms
Apoptosis
