实用肿瘤学杂志 ›› 2010, Vol. 24 ›› Issue (5): 401-403.doi: 10.3969/j.issn.1002-3070.2010.05.001

• 论著 •    下一篇

绿色荧光蛋白pAcGFP标记的人乳腺癌细胞株的建立

杨悦1, 王丽萍2, 杨艳梅1, 张春艳1, 马玉彦1, 蔡会龙1, 任丽君1   

  1. 1.哈尔滨医科大学肿瘤防治研究所(哈尔滨 150081);
    2.武警黑龙江总队医院病理科
  • 收稿日期:2010-05-06 出版日期:2010-10-28 发布日期:2015-01-24
  • 通讯作者: 杨艳梅,E-mail:yangym0916@163.com
  • 作者简介:杨悦,男,(1979-),本科,助理研究员,从事肿瘤动物模型的构建研究
  • 基金资助:
    黑龙江省科技厅科技计划项目(PC07S012)

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

YANG Yue1, WANG Liping2, YANG Yanmei1, ZHANG Chunyan1, MA Yuyan1, CAI Huilong1, REN Lijun1   

  1. 1.Harbin Medical University Cancer Research Institute,Harbin 150081;
    2.CAPF Heilongjing Hospital
  • Received:2010-05-06 Online:2010-10-28 Published:2015-01-24

摘要: 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。

Abstract: Objective To establish a human breast cancer cell line with green fluorescent protein(pAcGFP)gene.Methods Human breast cancer cell lines MCF-7 were transfected with pAcGFP-C1 by Fugene HD Transfection Reagent.The cells which express the pAcGFP gene were selected by geneticin(G418).The expression of pAcGFP 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 untransfected ones.Results A human breast cancer cell line with pAcGFP was established,named MCF7-pAcGFP.The transfection rate was 85%.The growth curve and invasive power of the transfected cells were similar to untransfected ones(P>0.05).Conclusion The MCF7-pAcGFP cells could stably express AcGFP,and pAcGFP transfection does not change the biological behavior of the cells.

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