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目的 :研究建立稳定高表达绿色荧光蛋白 (GFP)并能连续传代培养的肝癌细胞株。方法 :将携带GFPcDNA的重组逆转录病毒载体转染肝癌细胞株 772 1,经过G418筛选和克隆化培养 ,用荧光显微镜检测癌细胞荧光表达。结果 :转染GFPcDNA的癌细胞 5d后大部分死亡 ,荧光显微镜下有散在或小团状的点状荧光。继续用G418筛选及克隆化培养 6 5d后第 2 8培养孔的癌细胞几乎均见荧光表达并可连续稳定传至 15代以上的培养。结论 :GFP是较好的细胞报告基团 ,肝癌 772 1 GFP细胞株的建立有助于观察和了解肿瘤侵袭和转移发生及其规律。
Objective : To study the establishment of stable and highly expressed green fluorescent protein (GFP) and continuous subculture of hepatoma cell lines. METHODS: The recombinant retroviral vector carrying GFP cDNA was transfected into hepatoma cell line 772 1, screened by G418 and cloned and cultured. The fluorescence expression of cancer cells was detected by fluorescence microscopy. Results: Most of the cancer cells transfected with GFP cDNA died after 5 days. There were scattered or small clusters of dot-like fluorescence under fluorescent microscope. After continuous selection with G418 and clone culture, the cancer cells in the 2nd 8th culture wells were almost uniformly expressed in fluorescence and could be continuously transferred to cultures of more than 15 passages. Conclusion : GFP is a good cell reporter group. The establishment of hepatoma 772 1 GFP cell line is helpful to observe and understand the occurrence and regulation of tumor invasion and metastasis.