绿色荧光蛋白基因标记的肺癌细胞株的建立

Establishment of a lung cancer cell line marked by green fluorescent protein

目的建立能稳定高表达增强型绿色荧光蛋白(EGFP)的肺癌细胞株,为研究肺癌的浸润与转移机制打下基础。方法利用逆转录病毒将EGFP导入人高转移肺癌细胞株95D,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EGFP基因在癌细胞体外的表达,绘制细胞生长曲线,检测细胞贴壁率等,观察稳定表达EGFP的癌细胞的细胞生物学行为有无改变。结果携带EGFP的逆转录病毒能有效地感染95D细胞,筛选出的细胞能稳定、高效、持久地表达EGFP,与未感染的细胞比较,他们的生物学特性未改变。结论人高转移肺癌细胞株95D/GFP-1的建立为研究肺癌侵袭和转移的发生机制提供了理想的细胞株。

Objective In order to observe the infiltration and metastasis of the tumor,establish a high metastatic lung cancer cell line expressing enhanced green fluorescent protein （EGFP）. Methods 95D cell line was infected by the retrovirus which mediating expression of an enhanced green fluorescent protein gene and selected by G418, The expression of EGFP of the infected cells was observed by fluorescence microscopy and detected by flow cytometry. Multiple biological behaviors, such as the growth curve and cell adherent rate were compared between the infected and uninfected cells. Results 95D cell line could be infected efficiently by retrovirus. The selected cells could express the EGFP stably, efficiently and persistently. There was no significant difference between infected cells and uninfected cells in their biological behaviors. Conclusion The establishment of 95D/GFP-1 can offer a useful cell line to investigate the mechanisms of tumor＇s infiltration and metastasis.