肝癌细胞绿色荧光蛋白基因的转染与克隆化培养

Transfection by green fluorescent protein gene and cloned culture of cell line of hepatocellular carcinoma

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。

Purpose:To study the establishment of stable,high level green fluorescent protein (GFP) expressing cell line of hepatocellular carcinoma(HCC) and pass on its culture contiuously.Methods:7721 cell line of HCC was transfected by retrovirus vector with integrated GFP cDNA. GFP expressing cancer cells were detected by fluorescent microscopy through G418 selecting and cloned culture.Results:Most of the cancer cells were dead 5 days after being transfected by GFP, scattered or clustered green fluorescence can be seen by microscopy. In culture for 65 days,almost all of the clone 28 cells expressed high intensity GFP fluorescence and stability.Conclusions:7721 GFP cell line can provide a basis for establishing an ideal animal model for research of tumor invasion and metastasis.