Anti-hepatoma Effect of DC2.4 Cells Transfected with Tumor-Associated Antigen Cdc25C In Vitro  

作　　者：Chun-mei LI Yan-fei LI Lin TIAN Qi-hui ZHANG Fang-yuan ZHENG Fa-rong MO 

机构地区：[1]School of Basic Medical Science,Guangxi Medical University,Nanning 530021,China [2]Laboratory of Hepatobiliary and Pancreatic Surgery,Affiliated Hospital of Guilin Medical University,Guilin 541001,China [3]Guangxi Colleges and Universities Key Laboratory of Human Development and Disease Research,Guangxi Medical University,Nanning 530021,China

出　　处：《Current Medical Science》2022年第3期491-497,共7页当代医学科学（英文）

基　　金：the National Natural Science Foundation of China(No.81160264);the Natural Science Foundation of Guangxi Province(No.2016GXNSFAA380267 and No.2018GXNSFAA281071);the Guangxi First-class Discipline Project for Basic Medicine Sciences(No.GXFCDP-BMS-2018);the Research Enhancement Project for Junior Faculty in Higher Education Institutes of Guangxi(No.2018KY0419).

摘　　要：Objective Cell division cyclin 25 homolog C(Cdc25C)is a tumor-associated antigen candidate gene,and this may be used as an effective target in cancer treatment.The present study aims to evaluate the lysis effect of cytotoxic T lymphocytes(CTLs)induced by dendritic cell line DC2.4 overexpressing Cdc25C,and the feasibility of Cdc25C as a component in hepatoma immunotherapy.Methods The mouse Cdc25C gene was ligated into a lentiviral vector,and transfected into DC2.4 cells.The DC2.4 cell phenotype and cytokine secretion were determined by flow cytometry and ELISA,respectively.CD8^(+)T cells were sorted from the spleens of C57BL/6 mice using a magnetic bead sorting kit obtained from Miltenyi Biotech,Germany,and co-cultured with DC2.4 cells for one week as effector cells.Then,IL-2,granzyme B and perforin were detected in the CTL culture medium by ELISA.Next,time-resolved fluorescence immunoassay was used to detect the immune killing effect of Cdc25C-specific CTLs on target cells.Meanwhile,the effect of blocking MHC-I sites on target cells with a monoclonal anti-MHC-I antibody was evaluated.Results The results revealed that Cdc25C could be stably overexpressed in DC2.4 cells by LV-Cdc25C infection.DC2.4 cells transfected with LV-Cdc25C secreted more IL-6,IL-12,TNF-αand IFN-γ,and had higher expression levels of CD40,CD86,CCR7 and MHC-II than unaltered DC2.4 cells.The elevated Cdc25C in dendritic cells also further increased the secretion of IL-2,granzyme B and perforin to elicit Cdc25C-specific CTLs,and induced the higher cytotoxicity in Hepa1-6 cell lines(P<0.05),but this had no effect on the target cells when MHC-I monoclonal antibodies were blocked.Conclusion DC2.4 cells transfected with LV-Cdc25C can induce specific CTLs,and result in a strong cellular immune response.The dendritic cells that overexpress Cdc25C may be useful for hepatoma immunotherapy.

关 键 词：dendritic cells cell division cyclin 25 homolog C cytotoxic T lymphocytes hepatocellular carcinoma anti-hepatoma 

分 类 号：R73[医药卫生—肿瘤]