糖化多聚赖氨酸偶联MIP-3α—FL对小鼠肝癌免疫微环境的调节作用

The effect of glycated polylysine-coupled MIP-3a-FL on the immune microenvironment of mouse liver cancer

目的探讨重组糖化多聚赖氨酸偶联MIP-3α-FL双基因靶向表达载体在肝癌免疫微环境中树突状细胞（DC）的调节作用。方法采用MIP-3α—FL重组质粒（recombinant plasmid of MIP-3α-FL，shMIP-3α—FL）转染H22肝癌细胞，Westernblot和ELISA分析载体表达；重组糖化多聚赖氨酸偶联双基因靶向表达载体，经尾静脉注入肝癌模型小鼠，比较各组小鼠生存时间与肿瘤性浸润DCs的数量和表型。结果Western blot和ELISA证实，MIP-3α-FL基因转染后H22细胞MIP-3α和FL分泌明显增加。荷肝癌小鼠尾静脉注射重组糖化多聚赖氨酸偶联MIP-3α—FL双基因靶向表达载体后，实验组小鼠存活时间比对照组明显延长，肿瘤缩小。流式细胞术检查显示，实验组小鼠肿瘤性浸润DCs数量明显增加。实验组小鼠肿瘤浸润性树突状细胞表型CD80和CD86表达明显高于对照组。结论MIP-3α和FL共同作用可以明显促进DC集聚和成熟．偶联糖化多聚赖氨酸载体提高靶向作用精度，增强肿瘤免疫微环境中DC的抗原提呈作用。

Objective To study the regulation of dendritic cells by recombinant glycated polylysine-coupled MIP-3α-FL double-gene targeting expression vector in liver cancer immune microenvironment. Methods H22 hepatocarcinoma cells were transfected with recombinant plasmid of MIP-3α-FL （shMIP-3α- FL） and injected into hepatoma model mice. The survival time, tumor size were compared. Flow cytometry was used to measure the number and phenotype of tumor infiltrating DCs. Results Western blot and ELISA demonstrated that the secretion of MIP-3α and FL in H22 cells was significantly increased after transfcction with MIP-3α-FL . The survival time of the mice in the experimental group was significantly prolonged, the tumor size decreased. Flow cytometry showed that the number of tumor-infiltrating DCs in the experimental group was significantly higher than that in the control group ; the expression of CD80 and CD86 in the infiltrating dendritic ceils （TIDCs） was significantly higher than that of the control group. Conclusions The co-action of MIP-3cx and FL can significantly promote DC accumulation, maturation, and conjugate glycosylated polylysine carriers increase the precision of targeting and enhance the antigen presentation of the DCs.