共表达MAGE-1与EGFP基因的小鼠黑色素瘤细胞系的建立

Establishment of mouse melanoma B16 cell lines co-expressing MAGE-1 and EGFP

目的 :构建黑色素瘤抗原 1(MAGE 1)基因的真核表达载体 ,并在小鼠黑色素瘤B16细胞中进行表达。方法 :采用分子生物学的手段 ,构建了MAGE 1与增强型绿色荧光蛋白 (EGFP)基因共表达的质粒pIRES2 EGFP MAGE 1,通过脂质体以共表达质粒转染B16细胞 ,用荧光显微镜检测细胞中EGFP的表达 ,用免疫组化染色法检测细胞中MAGE 1的表达。结果 :成功地构建了真核表达载体pIRES2 EGFP MAGE 1。以其转染B16细胞后 ,经荧光显微镜及免疫组化染色法检测 ,可见细胞内有EGFP及MAGE 1的表达。结论 :成功地建立了可共表达MAGE 1与EGFP基因的B16细胞 ,为MAGE 1在肿瘤免疫治疗中的应用奠定了研究基础。

AIM: To construct the melanoma antigen-1(MAGE-1) eukaryotic expression plasmid and express MAGE-1 in mouse melanoma B16 cells. METHODS: The MAGE-1 gene was amplified by PCR and cloned into the eukaryotic expressi on vector pIRES2-EGFP to construct the pIRES2-EGFP-MAGE-1 plasmid. The plasm id was transfected into the B16 cells. The EGFP expression was detected under f luoroscent microscope and the MAGE-1 expression was detected by immunohistoche mistry staining. RESULTS: The eukaryotic expression vector pIRE S2-EGFP-MAGE-1 was constructed and transfected successfully into B16 cells, a nd the EGFP and MAGE-1 genes were co-expressed in the B16 cells. CONCL USION: A mouse melanoma cell line B16 co-expressing MAGE-1 and EGFP genes has been established successfully, which lays the foundation for the rese arch on application of MAGE-1 in the tumor immunotherapy.