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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。
As an important hematopoietic regulator, IL-3 not only acts on immature hematopoietic progenitor cells but also on mature immune cells. To study the immunoregulatory function of IL-3 and its potential anti-tumor effect, we established an autocrine IL-3 tumor cell using gene transfection technology. First, we constructed the IL-3 expression vector BMGNeo-mIL-3, which was then transfected into B16 mouse melanoma cells. A high-IL-3 expressing clone (806 U/ml) was obtained through G418 resistance screening and limited dilution. The expression of IL-3 in cells was confirmed by Northern hybridization. Studies on in vitro growth characteristics of this cloned strain revealed that although both wild-type B16 cells and B16-Neo cells transfected with the control plasmid BMGNeo do not express IL-3, the in vitro growth ability of B16 cells expressing IL-3 after introduction into the gene and No significant change. This laid the foundation for further study of the immune regulatory function of IL-3 and the anti-tumor effect in vivo.