基础医学与临床 ›› 2008, Vol. 28 ›› Issue (6): 579-582.

• 研究论文 • 上一篇    下一篇

在癌细胞中下调皮质肌动蛋白使细胞内吞作用减弱

陈莉 朱建伟 Zhan Xi   

  1. 江苏省南通大学医学院病理教研室 江苏省南通大学附属医院 UMB Greenebaum 肿瘤中心
  • 收稿日期:2007-04-19 修回日期:2005-06-19 出版日期:2008-06-25 发布日期:2008-06-25
  • 通讯作者: 朱建伟

Attenuated endocytosis function by down-regulation of cortactin protein in cancer cells

Li CHEN, Jian-wei ZHU, Xi ZHAN   

  1. Dept.of Pathologym Nantong University
  • Received:2007-04-19 Revised:2005-06-19 Online:2008-06-25 Published:2008-06-25
  • Contact: Jian-wei ZHU,

摘要: 目的 探讨皮质肌动蛋白(Cortactin,Cort)对高表达Cort的乳腺癌细胞系(MDA-MB-231细胞)细胞内吞作用的影响。方法 在MDA-MB-231细胞中使用CortsiRNA干预技术和"生物搬运法"系统将抗Cort免疫抗体(Cort多克隆抗体001或单克隆Cort抗体4F11)引入MDA-MB-231细胞,进行体内干预Cort功能。根据捕获ELISA法分析转铁蛋白摄入,免疫印迹分析细胞中Cort敲除的程度和免疫荧光直接检测评价Cort在转铁蛋白吸收中的作用。结果 免疫印迹分析在CortsiRNA处理过的细胞质中未测出Cort蛋白;用荧光标记评价转铁蛋白密度水平,在CortsiRNA处理的细胞中要低于含相对高水平Cort的对照组细胞;在接受CortsiRNA处理后仅50%的MDA-MB-231细胞中有转铁蛋白摄入。在MDA-MB-231细胞中引入001或4F11抗体,其细胞的内吞作用减弱,导致明显的转铁蛋白吸收减少30%~60%。结论 干扰Cort的功能或直接使用抗Cort抗体进行干预都能抑制细胞内吞作用。Cort是细胞内吞作用中必不可少的成分,具有重要作用。

关键词: 皮质肌动蛋白(Cort), 乳腺癌细胞株, siRNA, 转铁蛋白, 细胞内吞作用

Abstract: Objective To study cortactin function in cancer cell endocytosis. Methods We applied cortactin siRNA interference to MDA-Mb-231, a human breast adenocarcinoma cell line, in which cortactin is over-expressed, and introduced anti-cortactin immunoreagents into the cells using the BioPorter system to interfere with cortactin function in vivo. Capture -ELISA assay was used to measure transferrin uptake, immunoblot assay to assess the effect of cortactin knock-down, and immunoflurescence microscopy to examine the effect of cortactin down-regulation on transferrin uptake. Results Interference of cortactin function in cells led to the impairment of transferrin endocytosis. Transferrin fluorescent intensity in cytoplasm in cortactin siRNA treated-cells was apparently reduced in comparison to mock-treated cells. Less than 50% of cells subjected to cortactin siRNA treatment had normal transferrin uptake. Endocytosis in MDA-Mb-231 cells introduced with cortactin antibodies was impaired as well, showing a 30 ~ 60% reduction in transferrin uptake. Conclusions This study implies that cortactin, an actin-binding protein, plays an essential role in cell endocytosis

Key words: Cortactin, Cancer cell, Endocytosis, siRNA Interference