mRNA DC疫苗对肝癌细胞增殖周期影响的实验研究

Inhibitory effects of total mRNA-dendritic cell vaccine on hepatocellular carcinoma cell proliferation cycle

目的探讨mRNADC疫苗对肝癌细胞增殖周期的影响，为mRNA疫苗免疫治疗肝癌提供新的实验依据和理论基础。方法采用粒／巨噬细胞集落刺激因子（GM—CSF）和白介素-4（IL-4）联合培养、扩增鼠骨髓来源DC，以阳离子脂质体转染Hepal-6mRNA入DC，制备mRNADC疫苗，流式细胞术检测疫苗对体外培养的肝癌细胞周期以及体内种植瘤组织细胞周期的影响；免疫组织化学检测肿瘤细胞增殖核抗原（PCNA）。结果mRNA DC疫苗处理后的肝癌细胞周期比例为G0／G1间期（75．09±3．41）％，S间期（16．93±1．57）％，G2/M间期（7．98±1．83）％，与对照组比较，差别有显著性意义（P〈0.05）；荷瘤鼠疫苗组肿瘤细胞增殖指数为（24．91±3．41），与对照组比较，差别有统计学意义（P〈0．05）；PCNA实验组阳性表达率为（27．7±10．3）％，与对照组比较，差异有显著性意义（P〈0．05）。结论mRNA DC疫苗可影响肿瘤细胞的分裂周期，减缓细胞的生长，显著抑制肿瘤细胞增殖。

Objective To study the inhibitory effects of total mRNA-dendritic cell vaccine on hepatocellular carcinoma cell proliferation cycle. Methods DC isolated from mice marrow were incubated with recombinant mouse granulocyte macrophage colony-stimulating factor and mouse IL-4 and transfected with Hepal-6 total RNA by eathodolyte lipid. The inhibitory effects of total mRNA-dendritic cell vaccine on hepato cellular carcinoma cell proliferation cycle in vivo and in vitro were detected by flow cytometry. Immunohisto chemical staining was used to investigate the proliferating cell nuclear antigen. Result Treated with the mR NA vaccine in vitro, the percentages of G0/G1 ,S, G2/M cells of tumor cells were（75.09±3.41）%, （16.93 ± 1.57）%, （7.98±1.83）% respectively. There was significant difference in the percentages of tumor cells in different cell cycles between the experimental group and the control groups （P〈0.05）. The proliferative index of tumor cells treated with the mRNA vaccine was （24.91±3.41）, which was significantly lower than those in the control groups. The positive rate of the PCNA expression was（27.7 ± 10.3）% in the experiment group, which was significantly lower than that in the control groups （P〈0.05）. Conclusion mRNA DC vaccine can affect cell cycle of the tumor cells to inhibit tumor cell proliferation and growth.