mRNA高通量测序的树突状细胞原位胰腺癌细胞瘤苗抑制胰腺癌细胞的机制  

The mechanism of inhibiting the growth of pancreatic cancer cells by dendritic cell-biopsy xenograft of pancreatic carcinoma line-3 fusion vaccine by mRNA high-throughput sequencing

作　　者：徐亚晨 何志军 李凯 谢书海 孙早喜[1] 黄海溶 Xu Yachen;He Zhijun;Li Kai;Xie Shuhai;Sun Zaoxi;Huang Hairong(Department of General Surgery,the First Affiliated Hospital of Hainan Medical University,Haikou 570102,China;Department of Epidemiology,School of Public Health of Hainan Medical University,Haikou 571199,China)

机构地区：[1]海南医学院第一附属医院肝胆外科,海口570102 [2]海南医学院公共卫生学院流行病学教研室,海口571199

出　　处：《中华实验外科杂志》2020年第4期670-672,共3页Chinese Journal of Experimental Surgery

基　　金：海南省重点研发项目(2016116);海南省应用技术研发与应用推广专项(2015081)。

摘　　要：目的探讨mRNA高通量测序的树突状细胞(DC)原位胰腺癌细胞(BxPC-3)瘤苗(DC-BxPC-3)抗胰腺癌的机制。方法 2018年12月至2019年12月取新鲜外周静脉血(红十字会),胰腺癌BxPC-3细胞购自上海生命科学研究院。以淋巴细胞分离液分离50 ml外周静脉血,贴壁培养获得DCs和去DCs的系统免疫效应细胞(SIECs),并诱导增殖。DCs致敏时,BxPC-3∶DCs=1∶1;抑制与凋亡实验时,DCs∶SIECs∶靶细胞=1∶20∶2。以mRNA高通量测序,细胞计数试剂盒(CCK-8)、膜联蛋白V-异硫氰酸荧光素(Annexin V-FITC)、碘化丙锭(PI)进行检测。组间比较采用LSD或Dunnett T3检验。结果 CCK-8实验显示,致敏DCs组、DCs-BxPC-3瘤苗组的BxPC-3生存率分别为56.7%、23.2%,两组比较差异均有统计学意义(t=11.160,P<0.05)。凋亡实验显示,致敏DCs组、DCs-BxPC3瘤苗组对BxPC-3的凋亡率分别为(23.12±1.05)%、(80.86±5.09)%,组间比较差异有统计学意义(t=19.240,P<0.05)。差异mRNA的基因本体论(GO)分析显示融合瘤苗组的催化活性、分子信号传导活性、免疫系统、代谢过程、生物调节等功能富集;京都基因与基因组百科全书(KEGG)结果显示差异基因主要富集于干扰肿瘤细胞氨基酸代谢与脂质转运。结论 DC瘤苗诱导免疫效应细胞抗胰腺癌效能优于肿瘤裂解物致敏DC。Objective To probe the mechanism of anti-pancreatic cancer of dendritic cell(DC)-biopsy xenograft of pancreatic carcinoma line-3(BxPC-3)vaccine by high-throughput sequencing of mRNA.Methods A total of50 ml of peripheral venous blood was separated by lymphocyte separation fluid,and DCs and systematic immune effector cells(SIECs)were obtained by adherent culture,and then proliferation of DCs was induced.When DCs were sensitized,BxPC-3∶DCs=1∶1;when the experiments on the inhibition and apoptosis were done in BxPC-3,DCs∶SIECs∶target cells=1∶20∶2.The mRNA high-energy sequencing,cell counting kit-8(CCK-8),Annexin V-fluoresceine isothiocyanate(FITC),and propidium iodide(PI)methods were used for detection.Results The CCK-8 assay showed that the survival rate of BxPC-3 in the sensitized DCs group and DCs-BxPC-3 fusion vaccine group was 56.7%and 23.2%respectively(Dunnett’13 test,t=11.160,P<0.05).Apoptosis experiments showed that the sensitized DCs group and DCs-BxPC-3 tumor vaccine group had an apoptosis rate of(23.12±1.05)%and(80.86±5.09)%for BxPC-3,and the difference between two groups was statistically significant(t=19.240,P<0.05).Gene ontology(GO)analysis of differential mRNA showed that the tumor vaccine group was enriched in functions such as catalytic activity,molecular signaling activity,immune system,metabolic process,and biological regulation.The kyoto encyclopedia of genes and genomes(KEGG)results showed that differential genes were mainly enriched in interfering with tumor cell amino acid metabolism and lipid transport.Conclusion The antitumor effect of DC tumor vaccine to induce immune effector cells is better than that of tumor lysate-sensitized DC.

关 键 词：树突状细胞 胰腺癌 瘤苗 基因本体论 

分 类 号：R735.9[医药卫生—肿瘤]