表达β-半乳糖苷酶的gal B16肿瘤模型的建立及其在肿瘤DNA疫苗研究中的应用

ESTABLISHMENT OF MOUSE MELANOMA MODEL EXPRESSING β-GALACTOSIDASE AND ITS APPLICATION IN THE RESEARCH OF DNA VACCINES AGAINST TUMOR

本文工作的目的是建立以β-半乳糖苷酶为标志性抗原的小鼠黑色素瘤模型,并进行肿瘤免疫的研究。我们首先在pcDNA3质粒中引入一个β-半乳糖苷酶编码基因从而建立转染质粒p3gal。p3gal转染小鼠黑色素瘤细胞B16后,再通过G418筛选及X-Gal细胞染色得到表达β-半乳糖苷酶的galB16细胞株。接着用该细胞株成功地在C57小鼠上建立了表达β-半乳糖苷酶的galB16肿瘤模型。并在此模型上观察了β-半乳糖苷酶编码基因作为DNA疫苗抑制galB16肿瘤生长的作用。

We established a mouse melanoma model expressing β-galactosidase for the study of tumor immunotherapy.The recombinant vector p3gal was constructed by inserting a β-galactosi- dase gene into the MCS of plasmid pcDNA3.The vector then transfected the B16 cells.Through selection with 500μg/ml G418 and in situ X-Gal staining,the melanoma cell line galB16,stably expressing β-galactosidase was obtained.The melanoma model was successfully established after inoculation in mouse with galB16 cells.In situ X-Gal staining showed that the tumor ceils ex- pressed β-galactosidase in vivo.With the model,we designed animal experiments for mouse tumor immunotherapy.Twenty mice were randomly assigned to four parallel groups.They received i.m. injection with saline,DNA vaccine p3gal(100μg/mouse),adjuvant CpG 1826(20μg/mouse),or p3gal+CpG 1826 respectively.Our result suggested that the DNA vaccine containing β-galactosidase gene could protect mice against the galB16 tumor challenge.In addition,when combining with the adjuvant CpG 1826,the effect was increased prominently.