稳定表达Trop-2基因的NIH3T3细胞系的构建和特性分析
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家自然科学基金资助(81101704);南京市科技局(201201090)


Construction and characterization of Trop-2 stable expression in NIH3T3 cells
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    目的:建立稳定表达人滋养层细胞表面抗原(Trop-2)NIH3T3细胞,分析过表达Trop-2对NIH3T3细胞的生长-增殖和侵袭特性的影响。方法:将Trop-2基因克隆到真核表达载体pcDNA3.1,转染NIH3T3细胞,通过G418筛选及RT-PCR鉴定获得稳定表达Trop-2的NIH3T3细胞(NIH3T3-Trop-2)。用MTS法检测NIH3T3-Trop-2细胞的增殖能力,软琼脂集落形成实验检测NIH3T3-Trop-2细胞的克隆形成能力,明胶酶谱法检测NIH3T3-Trop-2细胞的基质金属蛋白酶(MMP)-2和MMP-9的分泌及细胞划痕实验检测NIH3T3-Trop-2细胞的迁移能力。结果:稳定表达Trop-2的NIH3T3细胞在生长增殖-克隆形成及侵袭能力均较NIH3T3细胞强,细胞培养上清中的MMP-2和MMP-9增多。结论:Trop-2对细胞的增殖与迁移能力具有明显的促进作用。

    Abstract:

    Objective:To establish an NIH3T3 cell line to stable express human Trop-2 gene,and analysis the affection of Trop-2 on proliferation,migration and aggressiveness of NIH3T3-Trop-2 cell. Methods:The human Trop-2 gene was cloned into eukaryotic expression system pcDNA3.1 and transfected into NIH3T3 cells. The Trop-2 stable expression cells were selected by G418 and confirmed by RT-PCR. The cell proliferation,migration and aggressiveness were detected by MTS and wound healing assay,MMP-2/MMP-9 was analyzed. Results:The proliferation of NIN3T3-Trop-2 was higher than NIN3T3 cell. The wound healing assay results showed and that Trop-2 improved the cell migration,increased the number of foci generated(P < 0.05) and increased MMP-9 expression(P < 0.05) when compared to the NIH3T3 control group. Conclusion:These show that human Trop-2 is sufficient to improve the cell proliferation and induce the transformation of NIH3T3 cells.

    参考文献
    相似文献
    引证文献
引用本文

刘 玉,唐小军,曹 清,丁贵鹏,张慧林,王 欢,郑 峰,徐凛锋,林 红.稳定表达Trop-2基因的NIH3T3细胞系的构建和特性分析[J].南京医科大学学报(自然科学版),2013,(7):867-872

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2013-02-23
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2013-07-09
  • 出版日期:
通知关闭
郑重声明