人剪切修复基因着色性干皮病基因D对人脐静脉内皮细胞的促凋亡作用
DOI:
作者:
作者单位:

作者简介:

通讯作者:

基金项目:


The Promoting Effects of Xeroderma Pigmentosum D Gene on Proliferation of Human Umbilical Vein Endothelia 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
    摘要:

    目的探讨人剪切修复基因着色性干皮病基因D(XPD)对人脐静脉内皮细胞(HUVEC)的促凋亡作用。方法用脂质体转染法瞬时转染HUVEC,转染重组质粒pEGFP-N2/XPD和空载质粒pEGFP-N2,并用未转染的与重组质粒pEGFP-N2/XPD和空载质粒pEGFP-N2具有相同遗传背景和代数的HUVEC作为空白对照。实验分为3组:正常对照组、pEGFP-N2组和pEGFP-N2/XPD组。用荧光显微镜观察绿色荧光蛋白报告基因表达情况,用流式细胞仪检测细胞凋亡情况,用RT-PCR和Western Blot检测XPD、Bcl-2、Bax和wt-p53表达量的变化,用MTT法观察细胞增殖活力。结果在荧光显微镜下,可在转染了重组质粒pEGFP-N2/XPD或空载质粒pEGFP-N2的细胞中观察到绿色荧光,即转染成功;流式细胞仪结果显示,重组质粒pEGFP-N2/XPD的转染引起细胞凋亡增加(p<0.05或p<0.01);RT-PCR和Western Blot检测发现,重组质粒pEGFP-N2/XPD的转染使得XPD表达增高(p<0.05),同时使得Bcl-2表达降低,Bax和wt-p53表达增高(p<0.05或p<0.01);MTT结果显示,重组质粒pEGFP-N2/XPD的转染抑制细胞增殖活力(p<0.05)。结论XPD能促进HUVEC凋亡,下调XPD的表达,有望成为治疗动脉粥样硬化的一个新靶点。

    Abstract:

    AimTo investigate pro-apoptosis effects of xeroderma pigmentosum D(XPD) gene on human umbilical vein endothelia cells (HUVEC).MethodsRecombinant plasmid pEGFP-N2/XPD and vacant vector plasmid pEGFP-N2 were transient transfected into HUVEC by liposome 2000 method,with the same genetic background and algebra HUVEC as blank controls.The experiments were divided into three groups: control group, pEGFP-N2 group and pEGFP-N2/XPD group.The expression of green fluorescent protein was observed through fluorescence microscopy; the cell apoptosis rate was examined by flow cytometry; through RT-PCR and Western Blot, the expression levels of XPD,Bcl-2 and Bax and wt-p53 were detected ;the cell growth was detected by MTT.ResultsGreen fluorescences were observed in the cells transfected with pEGFP-N2/XPD or pEGFP-N2, indicating that the plasmids were transfected successfully.Flow cytometry results showed that overexpression of XPD increased the apoptosic rate of HUVEC (p<0.05 or p<0.01).RT-PCR results and Western Blot results showed that the transfection of pEGFP-N2/XPD increased the expression of XPD,Bax and wt-p53 (p<0.05), decreased the expression of Bcl-2 (p<0.05);MTT results showed that the transfection of pEGFP-N2/XPD inhibited the cell growth(p<0.05).ConclusionsXPD gene can promote HUVEC apoptosis.Therefore, down-regulating the expression of XPD gene is likely to be potential molecular target for treatment of atherosclerosis.

    参考文献
    相似文献
    引证文献
引用本文

张南,李菊香,丁浩,洪葵,吴清华,程晓曙.人剪切修复基因着色性干皮病基因D对人脐静脉内皮细胞的促凋亡作用[J].中国动脉硬化杂志,2012,20(5):445~450.

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
历史
  • 收稿日期:2011-05-06
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: