论文部分内容阅读
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。
Objective To establish and identify C26 cell line of mouse colon cancer stably transfected with mouse IL-17 gene. Methods The recombinant plasmid pcDNA3.1 was transfected into mouse colon cancer cell line C26 by lipofection. The cell line stably expressing IL-17 was screened by G418. The cells were observed under microscope The morphology, RTPCR and immunofluorescence were used to detect the expression of target genes and proteins. Scratch repair assay was used to detect the migration ability of cells. MTS assay was used to detect the proliferation of cells in vitro. RESULTS: One C26 cell stably expressing IL-17 was obtained. The high expression of IL-17 gene and protein in C26 / IL-17 cells demonstrated that IL-17 could express IL-17. The high expression of IL-17 enhanced the migration of C26 cells, Weaken the in vitro proliferation ability of this cell. Conclusion The murine colon cancer cell line stably transfected with IL-17 gene was successfully established.