稳定转染小鼠IL-17全长基因的结肠癌细胞株的建立及基因转染对C26细胞的影响

Construction of a transfected colon cancer cell line expressing mouse IL-17 and the effect of gene transfection on C26 cell

目的建立稳定转染小鼠IL-17基因全长的小鼠结肠癌C26细胞株并进行鉴定。方法脂质体法将携带小鼠IL-17基因全长的真核表达载体pc DNA3.1转染小鼠结肠癌细胞C26,经G418筛选出稳定表达IL-17的细胞株,镜下观察细胞形态,RTPCR法、免疫荧光法检测目的基因和蛋白的表达;划痕修复实验检测细胞的迁移能力,MTS法检测细胞体外增殖能力。结果获得1株稳定表达IL-17的C26细胞,C26/IL-17细胞高表达IL-17基因及蛋白,证明该细胞可表达IL-17;IL-17高表达可以增强C26细胞的迁移能力,减弱该细胞的体外增殖能力。结论成功建立了稳定转染IL-17基因的小鼠结肠癌细胞株。

This study designed to establish a stable mouse full-length IL-17 gene-transfected C26 cell line and identify its function. C26 mouse colon cancer cell line was transfected with IL-17 full length gene（inserted into pc DNA-3.1 vector） using lipofectamine 2000 and selected with G418. Then cellular morphology, target gene, and protein expressions of the cells were analyzed by RT-PCR and immunofluorescence; the migration and proliferation of the cells in vitro were detected. Data showed C26 cell line stably transfected with mice full length IL-17 was constructed. In conclusion, IL-17 gene could highly express in C26/IL-17 cell, suggesting that this cell line could express functional IL-17. So we have successfully established a mouse colon cancer cell line which is stably transfected with IL-17 gene.