【摘要】 目的建立能连续传代稳定高表达绿色荧光蛋白(GFP)的胃癌细胞株,探讨胃癌的生长与转移特征。方法利用脂质体将携带GFP-cDNA的真核表达载体质粒转染胃癌细胞株GC9811-P,经过G418筛选和克隆化培养,用荧光显微镜及流式细胞仪检测转染的EGFP基因在癌细胞体内外的表达,MTT比色法观察细胞生长,Westernblot检测转染细胞和未转染细胞肿瘤相关抗原的表达。结果携带EGFP的真核表达载体能有效地转染胃癌细胞GC9811-P,转染的细胞能稳定、高效和持久地表达EGFP,与未转染细胞比较,他们的生物学特性未改变(P>0.05)。在裸鼠皮下肿瘤中,EGFP也能稳定、高效的表达。结论胃癌GC9811P-GFP的建立为研究肿瘤侵袭和转移的发生机制提供了理想的细胞株。更多还原

【Abstract】 Objective In order to observe the growth and metastasis of the tumor directely,we established a gastric cancer cell line expressing green fluorescent protein(GFP). Methods GC9811-P cell line was transfected by the eukaryotic expressive plasmid of GFP using liposome dosper, GFP-expressing cancer cell line selected by means of G418, fluorescence microscope and flow cytometer.Multiple biological behaviors of transduced cells such as the proliferating potential and the expression of various antigens were comparatively analyzed between untrtransfected and transfected in vitro. Results The eukaryotic expressive plasmid of EGFP was effectively transferred into GC9811-P cells. GC9811-P cells with the EGFP gene displayed stable and long-term EGFP expression . There was no significant difference between transfected cells and untransfected cells in expression of tumor antigens and proliferation rate in vitro,EGFP was stable and highly expressed in subcutaneous. Conclusion This well characterized cell line should be useful to investigate the mechanisms of tumor’s metastasis.更多还原