siRNA干扰沉默KLK12基因抑制胃癌细胞生长的研究
作者:
通讯作者:
作者单位:

作者简介:

曹晖 E-mailcaohuishcn@hotmail.com

基金项目:

上海市科学技术委员会科研计划项目(08411964200)。


Experimental study on silencing the KLK12 expression by siRNA to inhibit the growth of gastric cancer 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
  • |
  • 音频文件
  • |
  • 视频文件
    摘要:

    目的:利用RNA干扰(RNAi)技术,构建KLK12-siRNA表达载体,探讨其在体外对胃癌细胞生长的影响。
    方法:合成KLK12-siRNA,以脂质体法转染至胃癌MKN-45细胞中建立稳定细胞系,并分成3组,即空白对照组、阴性对照组和KLK12 siRNA组。采用实时定量PCR和Western blot方法观察KLK12 siRNA转染后胃癌细胞KLK12基因和相应蛋白表达的变化;用MTT法,细胞迁移和侵袭试剂盒分别检测转染细胞的增殖、迁移和侵袭能力的变化。
    结果:KLK12 siRNA组的KLK12 mRNA和蛋白表达水平较两个对照组明显下调(P<0.05)。KLK12基因沉默显著抑制胃癌细胞的增殖,并降低体外胃癌细胞的迁移能力。
    结论:KLK12-siRNA表达载体能抑制胃癌MKN-45细胞的增殖和迁移,为胃癌的基因治疗提供了有益的实验依据。

    Abstract:

    Objective:To determine the inhibitory effect of synthetic KLK12 short interfering RNA on the expression of KLK12 gene in human gastric cancer line MKN-45 cells, and to elucidate its effect on proliferation of MKN-45 cells in vitro.
    Methods:The constructed KLK12-siRNA plasmid was stably transfected into MKN-45 cells by lipofectamine,then, MKN-45 cells were divided into negative control group,blank control group and KLK12 siRNA group. KLK12 expression was determined by real time PCR and Western blot. The proliferation,migration and invasion of transfected MKN-45 cells were evaluated by MTT assay,cell Migration and Invasion Assay Combo Kit.
    Results:KLK12 siRNA significantly inhibited the expression of KLK12 in gastric cancer cells at both mRNA and protein level. The proliferation of MKN-45 cells in KLK12 siRNA group was significantly suppressed and the ability of migration was also significantly decreased in vitro.
    Conclusions:The KLK12-siRNA expression vector can effectively suppress the proliferation and migration of MKN-45 cells,which may provide a novel gene therapy for gastric cancer.

    参考文献
    相似文献
    引证文献
引用本文

沈志勇| 赵恩昊| 刘骅| 金鑫| 曹晖. siRNA干扰沉默KLK12基因抑制胃癌细胞生长的研究[J].中国普通外科杂志,2010,19(10):1071-1075.
DOI:10.7659/j. issn.1005-6947.2010.10.005

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
历史
  • 收稿日期:2010-05-25
  • 最后修改日期:2010-09-10
  • 录用日期:
  • 在线发布日期: 2010-10-15