高强度聚焦超声制备DC瘤苗及其对BALB/c小鼠肿瘤治疗作用的实验研究

目的用高强度聚焦超声(HIFU)制备肿瘤抗原致敏树突状细胞(DC),观察其对BALB/c小鼠肿瘤的治疗作用。方法用剂量为1000W/cm2×30 s的HIFU辐照体外培养的CT-26结肠癌细胞,获取肿瘤抗原,与体外培养扩增第5天的DC共孵育,制备DC瘤苗。正常BALB/c小鼠皮下接种活的CT-26结肠癌细胞,7天后分别经皮下注射HIFU辐照制备的DC瘤苗,一周后以相同剂量加强治疗。3周后处死老鼠,剥离肿瘤,测量肿瘤重量和肿瘤体积及祛瘤小鼠的净重,剩余观察小鼠的生存时间及生存率。并设立反复冻融法致敏DC组(冻融组),单纯的DC细胞组(单纯DC组),以及等量的无血清培养液组(阴性对照组),比较各组是否有差异。结果 HIFU组、冻融组与单纯DC组、阴性对照组比较3周时肿瘤重量、肿瘤体积、祛瘤小鼠净重、小鼠中位生存时间等差异均有显著性意义(P均<0.01或P均<0.05)。HIFU组与冻融组比较肿瘤重量、体积及小鼠中位生存时间差异亦有显著性意义(P<0.05)。结论 HIFU辐照法制备的DC瘤苗对小鼠肿瘤有一定的治疗作用,并优于反复冻融法制备的DC瘤苗。

A study of making dc tumor vaccine by high intensity focused ul trasound and observing its antitumor

Objective: To establish a method of expanding dendritic cells from mice bone marrow in vitro and identify it with morphological,immunological phenotype determination to investigate the feasibility of high intensity focused ultrasound in making tumour antigen and DCs vaccine,and to investigate the active antitumor immunoprotecitive effect after inoculating mice by the production.Methods: The cultured CT-26 tumor cells were irradiated by dose of 1000W/cm2×30 s,then tumor antigen was acquired,and this antigen was co-incubated with DC that had been cultivated for 5days in vitro so as to make DC tumor vaccine.The active immunoprotecitive effect of DC tumor vaccine in vivo was observed in the four groups established: the HIFU group(group A),the repeated freezed group(group B),the simple DC group(group C),and negative control group(group D).The normal BALB/c mice were inoculated CT-26 colon cancer cells(5×105DC cells one mouse) subcutaneously,and after seven days,the mice were vaccinated by the acquired DC tumor vaccine(5×105) cells,with one mouse at one-time,and seven days later,the mice were vaccinated by the acquired DC tumor vaccine(5×105) cells with one mouse at one-time again.After 7 days the partial mice were executed to take out the tumor to survey tumor weight,the mouse dispel the lump net weight and the tumor volume,the surplus mouse was used to observe the medium survival time,the survival rate and whether there is differences in groups.Results: In the comparison between the HIFU group and freezing group and the negative group,there were remarkable differences in intumor weight and size at 20 days after seed(P<0.01 or P<0.05);In the comparison between the HIFU group and the freezing group,there were emarkable differences in tumor weight and size,mice medium survival time and survival rate.In the comparison of survival rates among three groups,difference was remarkable.These indicated that DC tumor vaccine derived by HIFU and repeated freezing may effectively inhibit the proliferation.Notable difference was also observed between the HIFU group and the freezing group,and more effective effect was seen in the HIFU group.Conclusion: Tumor vaccine prepared by HIFU have a certain therapeutic effect on mice tumors and may be better than tumor vaccines prepared by the repeated freezing.