基础医学与临床 ›› 2012, Vol. 32 ›› Issue (2): 211-216.

• 研究论文 • 上一篇    下一篇

外源EGFP表达对肿瘤细胞体外生物学行为的影响

张敏1,刘玉琴2,赵文晶1,李占稳1,冯海凉3,刘艳艳1   

  1. 1. 中国医学科学院基础医学研究所
    2. 中国医学科学院基础医学研究所;北京协和医学院基础学院
    3. 中国协和医科大学基础医学院,中国医学科学院基础所
  • 收稿日期:2011-11-08 修回日期:2011-12-21 出版日期:2012-02-05 发布日期:2012-01-12
  • 通讯作者: 刘玉琴 E-mail:ccc@pumc.edu.cn

Exogenous EGFP labeling affects Tumor Cells Biological Behavior

  • Received:2011-11-08 Revised:2011-12-21 Online:2012-02-05 Published:2012-01-12

摘要: 摘要:目的 探讨常用的可视化细胞标记技术--外源EGFP表达对肿瘤细胞生物学行为的影响。 方法 选择不同的携带EGFP的载体,利用脂质体转染或慢病毒感染技术,在不同肿瘤细胞系中强制表达外源EGFP,筛选稳定表达外源EGFP的细胞系,MTT法检测细胞体外增殖能力;细胞分析计数仪(CasyTT型)检测细胞大小的分布状况; Transwell体外迁移实验检测细胞体外侵袭能力。结果 成功建立了稳定表达外源EGFP的人结肠癌细胞系HCT116-GFP、HCT116-EGFP、COLO 320DM-EGFP和小鼠树突状细胞肉瘤细胞系DG6-EGFP。 外源EGFP表达后,HCT116-GFP细胞的增殖能力没有明显改变;而HCT116-EGFP细胞、COLO320DM-EGFP细胞和DG6-EGFP细胞的体外增殖能力明显降低。外源EGFP表达后,HCT116-EGFP细胞体积增大:平均直径分别为14.53 μm(HCT116)和18.28 μm(HCT116-EGFP);DG6-EGFP细胞体积无明显改变,平均直径分别为:16.43 μm(DG6)和16.58 μm(DG6-EGFP)。外源EGFP表达后,HCT116-EGFP细胞的体外侵袭能力明显降低,DG6-EGFP细胞体外侵袭能力无明显改变。结论 外源EGFP可视化标记细胞后,可能会影响肿瘤细胞的生物特性,利用这些模型进行科研时,需注意对相关指标的影响。

关键词: EGFP, 肿瘤, 细胞系, 增殖, 侵袭

Abstract: Abstract: Objective To study the influence of exogenous EGFP labeling on biological behavior of tumor Cells in vitro . Methods Using liposome transfection and lentivirus infection respectively, exogenous EGFP carried by different vectors was expressed in different tumor cell lines. Stable exogenous EGFP expressing cell lines were selected for further analysis. The growth was measured by MTT assay. The distribution of cell size was tested by Cell Counter(CasyTT). The invasion was measured by Boyden-chamber assay. Results Stably-expressing of exogenous EGFP human colon cancer cell line HCT116-GFP, HCT116-EGFP, COLO 320DM-EGFP and murine dendritic cell sarcoma cell line DG6-EGFP was successfully established. After expression of exogenous EGFP, the growth of HCT116-GFP cells was similar to that of HCT116 cells, while the growth of HCT116-EGFP cells, COLO320DM- EGFP cells and DG6-EGFPcells was significantly decreased. After expression of exogenous EGFP, the cell volume of HCT116-EGFP cells was bigger than that that of HCT116 cells, the cell volume of DG6-EGFP cells was similar to that of DG6 cells. After expression of exogenous EGFP, the invasion of HCT116-EGFPcells was significantly lower than that of HCT116 cells, the invasion of DG6-EGFP cells was similar to that of DG6 cells. Conclusion Exogenous EGFP labeling of tumor cells may or may not affect the biological characteristics of the tumor cells in vitro, indicating that before the use of these models, researchers should evaluate its effects on the relevant parameters.

Key words: EGFP, tumor, cell lines, proliferation, invasion

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