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目的:研究干扰素诱导的跨膜蛋白1(IFITM1)对结肠癌细胞SW480增殖和异质黏附的影响.方法:构建pEGFP-C3/IFITM1重组质粒,重组质粒转染结肠癌细胞株SW480,G418筛选获得亚克隆细胞株,用激光共聚焦扫描检查绿色荧光蛋白.生长曲线法检测细胞的增殖,细胞黏附实验检测细胞的黏附性.结果:pEGFP-C3/IFITM1重组质粒的测序序列与IFITM序列完全一致,重组质粒转染SW480细胞后,通过G418持续压力筛选获得亚克隆细胞株IFITM1/SW480.激光共聚焦显微镜检查结果显示表达的绿色荧光蛋白位于IF-ITM1/SW480的细胞膜上,提示转染后IFITM1正确表达.细胞生长曲线显示转染后的IFITM1/SW480细胞增殖活性比对照组增殖活性明显降低.IFITM1/SW480细胞黏附数30,60min分别为(3.93±0.08)×103和(6.10±0.21)×103,而对照组pEGFP-C3/SW480细胞黏附数30,60min分别为(4.61±0.25)×103,(7.17±0.22)×103,IFITM1/SW480细胞黏附数比对照组细胞黏附数显著降低(P<0.05).结论:IFITM1可抑制SW480细胞体外增殖和异质型黏附.
Objective: To investigate the effect of interferon-inducible transmembrane protein 1 (IFITM1) on proliferation and heterogeneous adhesion of colon cancer cell line SW480.Methods: The recombinant plasmid pEGFP-C3 / IFITM1 was constructed and transfected into colon cancer cell lines SW480 and G418 The subclone cell lines were obtained and the green fluorescent protein (GFP) was detected by confocal scanning laser scanning microscope.The growth curve was used to detect the cell proliferation and the cell adhesion assay was used to detect the cell adhesion.Results: The sequence of pEGFP-C3 / IFITM1 recombinant plasmid was identical with that of IFITM The recombinant plasmids were transfected into SW480 cells and then subcloned into IFITM1 / SW480 by continuous pressure screening with G418.Laser confocal microscopy showed that the expressed green fluorescent protein was located on the cell membrane of IF-ITM1 / SW480, indicating that IFITM1 .The results of cell growth curve showed that the proliferation activity of IFITM1 / SW480 cells after transfection was significantly lower than that of the control group (3.93 ± 0.08 × 103 and (6.10 ± 0.21) × 103, while the adhesion number of pEGFP-C3 / SW480 cells in control group was (4.61 ± 0.25) × 103 and (7.17 ± 0.22) × 103 respectively at 30 and 60min. The adhesion number of IFITM1 / SW480 cells was higher than that of control group Significantly lower (P <0.05) Conclusion:. IFITM1 inhibit proliferation of SW480 cells in vitro and idiosyncratic adhesion.