重组CCL2小鼠黑色素瘤B-16细胞的建立及生物学活性的鉴定  

Recombining murine CCL2 into melanoma B-16 cells and identifying their biological activity

作　　者：胡凯猛[1] 熊俊[1] 冀凯宏[1] 汤淑萍[1] 刘厚奇[1] 

机构地区：[1]第二军医大学组织学与胚胎学教研室,上海200433

出　　处：《解剖学杂志》2005年第4期397-399,F0002,共4页Chinese Journal of Anatomy

基　　金：国家自然科学基金(90208026)

摘　　要：目的:在小鼠黑色素瘤B-16细胞中克隆表达趋化因子CCL2,以进一步研究趋化因子与肿瘤的关系。方法:从小鼠腹腔巨噬细胞中提取总RNA,RT-PCR扩增CCL2 cDNA,将此片段重组于真核表达载体pcDNA3.0,脂质体FuGENE6转染鼠黑色素瘤B-16细胞,G-418筛选阳性克隆。结果:RT-PCR和免疫细胞化学鉴定转染B-16细胞中有较强CCL2的表达,体外趋化实验表明重组CCL2有生物学活性。结论:获得有趋化活性的重组CCL2的小鼠黑色素瘤B-16细胞。To clone chemokine CCL2 into murine melanoma B-16 cells and to investigate the relationship between cancer and chemokine. Methods： Total RNA was extracted from murine active macrophages in abdomen. CCL2 cDNA was amplified by RT-PCR and was cloned into pcDNA 3.0, and then the recombinant plasmid was transfected into melanoma B-16 cells. Results： The high production of CCL2 was certificated with RT-PCR and immunocytochemistry. And biological activity of CCL2 was detected with chemotaxis assay. Conclusion： The strains for high level CCL2 production melanoma B-16 cells have been developed, which can secret CCL2 with chemotaxis function.

关 键 词：小鼠 黑色素瘤 B-16细胞 生物学活性 重组单核细胞趋化蛋白1 

分 类 号：R73-3[医药卫生—肿瘤]