基础医学与临床 ›› 2011, Vol. 31 ›› Issue (9): 1000-1005.

• 研究论文 • 上一篇    下一篇

细丝蛋白A抑制人结肠癌SW480细胞体外侵袭转移能力

史建伟1,王贵英2,王士杰3,于跃明3,张娟3   

  1. 1. 河北大学附属医院
    2. 河北医科大学第四医院外二科
    3.
  • 收稿日期:2010-09-14 修回日期:2010-11-15 出版日期:2011-09-05 发布日期:2011-09-05
  • 通讯作者: 王贵英 E-mail:tizq12@vip.163.com
  • 基金资助:
    省教委基金;省卫生厅科研基金

Inhibitory effect of filamin A on invasion and metastasis of human colon carcinoma cell line SW480 in vitro

  • Received:2010-09-14 Revised:2010-11-15 Online:2011-09-05 Published:2011-09-05

摘要: 摘要 目的 研究细丝蛋白A(filamin A,FLNa)对人结肠癌SW480细胞侵袭转移行为的影响,并初步探讨其抑癌的机制。方法 将FLNa基因质粒载体pcDNA3.1/V5-His-TOPO/FLNa,以脂质体介导方式转染SW480细胞。经G418筛选,获得FLNa基因稳定表达的SW480 /FLNa细胞。逆转录聚合酶链反应(RT-PCR)和Western blot技术检测FLNa基因的导入情况;RT-PCR和Western blot检测了SW480细胞中基质金属蛋白酶-9(matrix metalloproteinases-9,MMP-9)的表达水平;MTT法检测细胞的增殖能力;划痕损伤实验、Transwell小室和Matrigel侵袭模型评价FLNa对细胞侵袭和转移能力的影响。结果 RT-PCR和Western blot证实FLNa的表达载体pcDNA3.1/V5-His -TOPO/FLNa在SW480细胞中获得稳定表达;SW480/FLNa细胞MMP-9的mRNA和蛋白表达水平低于野生型SW480细胞;划痕损伤实验、Transwell小室和Matrigel侵袭模型表明FLNa抑制SW480细胞的侵袭和转移。结论 FLNa能够抑制SW480细胞的侵袭和转移能力,其机制可能与降低MMP-9的表达有关。

关键词: 结肠肿瘤, 细丝蛋白A, 侵袭, 转移

Abstract: Abstract:Objective To explore the inhibitory effect of Filamin A(FLNa) on invasion and metastasis of human colon carcinoma cell line SW480 and investigate its mechanisms of the action.Methods Expressing vector pcDNA3.1/V5-His-TOPO/FLNa was transfected into SW480 cells by lipofectamine protocols.After G418 selection,the cells SW480/FLNa expressing FLNa stably were obtained.Expression of FLNa in tumor cells was confirmed by reverse transcription polymerase chain reaction(RT-PCR) and Western blot.RT-PCR assay and Western blot was also employed to detect the expression of matrix metalloproteinases-9(MMP-9).Cell proliferation in vitro were assessed by MTT assay.Inhibitory effect of filamin A on SW480 cells were observed by Wound healing assay,Transwell chamber experiment,and Matrigel invasion assay.Results The results of RT-PCR and Western blot demonstrated that the recombinant vector was correctly transfected into SW480 cells by liposome.The expression of MMP-9 mRNA and protein was lower in the SW480/FLNa cells than that in the wild SW480 cells;The inhibitory effect of FLNa on invasion and metastasis of SW480 cells were proved by wound healing assay,Transwell chamber experiment and matrigel invasion assay.Colusion FLNa has significant inhibitory effect on the invasion and metastasis of human colon carcinoma cell line SW480.The mechanism of action may be ralated with the suppressive effect of MMP-9.

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