人天冬氨酸-β-羟化酶树突状细胞疫苗对肝癌细胞系的体外作用研究  

Function of dendritic cells vaccines targeting aspartate-β-hydroxylase on hepatocellular carcinoma in vitro

作　　者：周玉姣 石果[2] 许晓蕾 罗雪 张苑玲 付婧婕 曾爱中[1] Zhou Yujiao;Shi Guo;Xu Xiaolei;Luo Xue;Zhang Yuanlin;Fu Jingjie;Zeng Aizhong(Department of Infectious Disease,The First Affiliated Hospital of Chongqing Medical University, Key Laboratory of Chongqing for Infectious Disease and Parasitic Disease;Department of General Surgery,The First Affiliated Hospital of Chongqing Medical University)

机构地区：[1]重庆医科大学附属第一医院感染科重庆市传染病寄生虫病学重点实验室,重庆400016 [2]重庆医科大学附属第一医院普外科,重庆400016

出　　处：《重庆医科大学学报》2019年第2期187-192,共6页Journal of Chongqing Medical University

摘　　要：目的:通过对携带人天冬氨酸-β-羟化酶(aspartate-β-hydroxylase,AAH)基因的树突状细胞杀伤肝癌细胞系的研究,探索可用于防治肝癌的树突状细胞疫苗。方法:重组腺病毒(Ad-AAH-IRES2-EGFP)感染C57BL/6小鼠骨髓未成熟树突状细胞(dendritic cells,DCs)制备携带AAH基因的DCs(AAH-DCs),与同体小鼠脾脏T淋巴细胞共培养后作为效应细胞,以肝癌细胞株HepG2和Huh7分别作为靶细胞,效应细胞分别和靶细胞共培养24 h,CCK-8法检测效应细胞对靶细胞增殖的影响;流式细胞术检测效应细胞对靶细胞凋亡的影响。将AAH-DCs连续3次皮下注射免疫C57BL/6小鼠,乳酸脱氢酶释放法检测免疫后小鼠脾脏T淋巴细胞对靶细胞的裂解效应。结果:AAH-DCs与T淋巴细胞共培养组靶细胞的增殖能力明显低于对照组(P=0.000),凋亡率明显高于对照组(P=0.000);实验组靶细胞的裂解率明显高于对照组(P=0.000)。结论:荷载AAH基因的树突状细胞疫苗能够诱导产生细胞毒性T淋巴细胞从而杀伤表达AAH的肝癌细胞。Objective:To explore the antitumor effect of dendritic cells(DCs)vaccine targeting aspartate-β-hydroxylase(AAH)gene against hepatocellular carcinoma(HCC)cells HepG2 and Huh7 for searching DCs vaccine for hepatocarcinoma. Methods:AAH-DCs vaccine was prepared in vitro via infecting DCs with recombinant adenovirus vector encoding AAH. T cells co-cultured with AAH-DCs vaccine were served as the effector cells and human HCC cell lines HepG2 and Huh7 were regarded as the target cells. The proliferation and apoptosis of target cells,after co-culturing with the effector cells for 24 h,were detected by CCK-8 and FCM analysis respectively. C57 BL/6 mice were immunized with AAH-DCs vaccines by three continuous subcutaneous injections,and the cytotoxic effect of T cells against target cells was observed by lactate dehydrogenase(LDH)release assay. Results:Compared with the control groups,the declined proliferation activity of target cells in AAH-DCs vaccine group was found by CCK-8,while increased apoptosis rate of target cells in AAH-DCs vaccine group was tested by FCM(P=0.000). Additionally,LDH release assay showed that the cytolysis effect of target cells in AAH-DCs vaccine group was higher than that in other control groups(P =0.000). Conclusion:DCs vaccines targeting AAH can induce the production of cytotoxicity T lymphocytes for fulfilling the killing of HCC cells expressing AAH gene.

关 键 词：肝细胞癌 树突状细胞 细胞毒性T淋巴细胞 天冬氨酸-β-羟化酶 

分 类 号：R392.11[医药卫生—免疫学] R735.7[医药卫生—基础医学]