绿色荧光蛋白pAcGFP标记的人乳腺癌细胞株的建立

Construction of human breast cancer cell line with green fluorescent protein pAcGFP

目的 构建绿色荧光蛋白（pAcGFP）标记的MCF-7细胞系.方法 应用Fugene HD Transfection Reagent转染MCF-7细胞.经G418筛选获得稳定表达pAcGFP的MCF-7细胞系,用倒置荧光显微镜检测pAcGFP的表达,用流式细胞仪检测pAcGFP转染率.通过生长曲线和体外侵袭实验,对转染和未转染pAcGFP细胞的生物学行为进行综合分析.结果 获得了稳定表达pAcGFP的乳腺癌细胞株MCF7-pAcGFP,转染率约为85%.生长曲线和体外侵袭力与未转染前相似（P＞0.05）.结论 MCF7-pAcGFP细胞株具备稳定表达AcGFP的能力,转染pAcGFP对MCF-7细胞的生长和转移特性并没有影响.

Objective To establish a human breast cancer cell line with green fluorescent protein （pAeGFP） gene. Methods Human breast cancer cell lines MCF - 7 were transfected with pAeGFP - C1 by Fugene HD Transfection Reagent. The cells which express the pAcGFP gene were selected by genetiein（G418）. The expression of pAeGFP were detected by inverted fluorescence microscope and flow cytometry, the growth curve and invasive power were observed to compare the biological behavior of the cells transfected with pAcGFP and untransfeeted ones. Results A human breast cancer cell line with pAeGFP was established, named MCF7 - pAcG- FP. The transfection rate was 85%. The growth curve and invasive power of the transfected cells were similar to untransfeeted ones （ P 〉 0.05 ）. Conclusion The MCF7 - pAeGFP cells could stably express AeGFP, and pAcGFP transfection does not change the biological behavior of the cells.