IL-3基因转染的黑色素瘤细胞株的建立及其体外生长性质的观察

Transfection of B16 Melanoma Cells with IL-3 Gene and Its in Vitro Proliferative Features

IL-3作为一种重要的造血调控因子,不仅作用于未成熟的造血前体细胞,还可作用于成熟的免疫细胞。为研究IL-3的免疫调节功能及其潜在的抗肿瘤作用,我们用基因转染技术建立了自分泌IL-3的肿瘤细胞。首先,我们构建了IL-3的表达载体BMGNeo-mIL-3,然后转染B16小鼠黑色素瘤细胞,通过G418抗性筛选和有限稀释获得高表达IL-3的克隆株(806U/ml),并经Northern杂交证实IL-3在细胞内的表达。对此克隆株的体外生长特性的研究发现,虽然野生型B16细胞和转染对照质粒BMGNeo的B16-Neo细胞均不表达IL-3,导入基因后表达IL-3的B16细胞的体外生长能力并无明显改变。这为进一步研究IL-3的免疫调节功能和体内的抗肿瘤作用打下了基础。

IL-3 has effects on a wide variety of cell types, including immature cells of the immune cells, and mature immune cells such as granulocytes. In the purpose of studying the immunoregulatory function of IL-3 and its potential role in cancer therapy, we established a IL-3-secreting tumor model using gene transfection to deliver locally IL-3 to tumor site. First, we constructed IL-3 expression vector BMGNeo-mIL-3, then transfected it into B16 murine melanoma cells. By G418 resistant screening and limiting dilution, we obtained a transfectant that expressed high levels of IL-3 (806U / ml) . The IL-3 expression of the transfectant was confirmed by Northern blot analyses. Although the wild-type B16 cells and the B16-Neo cells transfected with BMGNeo did not express IL-3, the IL-3 expression in B16 cells had no obvious effect on the in vitro proliferation of the transfectant. These results showed we had successfully established a IL-3-secreting tumor cell clone which would enable us to further study its in vivo tumorigenicity and immune function.