解剖学报

解剖学报 ›› 2014, Vol. ›› Issue (2): 221-227.doi: 10.3969/j.issn.0529-1356.2014.02.014

• 肿瘤生物学 • 上一篇    下一篇

姜黄素诱导人食管癌EC9706细胞凋亡过程中核磷蛋白的表达与定位变化

石松林1 陈兰英2 刘用金1 杨海波2 路锟1 杨玲1 李祺福1*   

  1. 1. 厦门大学医学院基础医学部，福建 厦门 361102; 2. 河南城建学院生命科学与工程学院，河南 平顶山 467036
  • 收稿日期:2013-05-14 修回日期:2013-06-21 出版日期:2014-04-06 发布日期:2014-04-06
  • 通讯作者: 李祺福 E-mail:chifulee@xmu.edu.cn
  • 基金资助:

    Nucleophosmin 蛋白修饰变化在肿瘤细胞增殖分化与凋亡调控中的作用机制研;HnRNP A2/B1 在乳腺癌细胞中的作用及分子机制;Prohibitin在肿瘤细胞增殖与分化调控中的作用机制研究

Expression and localization changes of nucleophosmin during apoptosis of human esophageal cancer EC9706 cells induced by curcumin

SHI Song-lin1 CHEN Lan-ying2 LIU Yong-jin1 YANG Hai-bo1 LU Kun1 YANG Ling1 LI Qi-fu 1*   

  1. 1.Department of Basic Medicine, Medical College of Xiamen University, Fujian Xiamen 361102, China;2. College of Life Science and Engineering, He’nan University of Urban Construction, He’nan Pingdingshan 467036, China
  • Received:2013-05-14 Revised:2013-06-21 Online:2014-04-06 Published:2014-04-06
  • Contact: LI Qi-fu E-mail:chifulee@xmu.edu.cn

摘要：

目的 探讨核磷蛋白NPM在癌细胞诱导凋亡过程中在细胞内、细胞核基质上的定位与表达变化，以及NPM与凋亡调控相关蛋白的关系，探索其在凋亡调控中的作用。 方法 在姜黄素诱导人食管癌EC9706细胞凋亡的基础上，以亚细胞蛋白质组学方法分析NPM在核基质中的存在与变化，并以免疫印迹法杂交实验进行确证；激光扫描共焦显微镜观察NPM在EC9706细胞凋亡过程中的定位与变化，以及NPM与Bax、Bcl-2等基因产物的共定位关系。 结果 NPM存在于EC9706细胞核基质蛋白组分中,并在姜黄素处理后表达下调。NPM在EC9706细胞凋亡过程中发生显著的胞质-核之间的穿梭定位变化，并与Bax、Bcl-2等蛋白具有共定位关系，且共定位区域发生了变化。 结论 NPM是一种核基质结合蛋白，在EC9706细胞凋亡中的表达与定位变化，及其与凋亡调控蛋白的共定位关系提示，它在EC9706细胞凋亡调控中具有重要作用。

Abstract:

bjective To explore the expression and localization change of nucleophosmin(NPM) in the nuclear matrix of cancer cells and its relationships with apoptosis regulating proteins during induced apoptosis, and to discuss its regulating functions in apoptosis. Methods Curcumin was used to induce apoptosis of human esophageal cancer EC9706 cells. By using subcellular proteomics technology we analyzed the presence and expression change of NPM in the nuclear matrix. We performed a Western blotting analysis to confirm the change. Subsequently, we examined the localization change of NPM and its co-localization with apoptotic gene products such as Bax and Bcl-2 through laser scanning confocal microscopy. Results The results of this study showed that NPM existed in the nuclear matrix of EC9706 cells with a remarkable down-regulation after curcumin treatment. We demonstrated that NPM exhibited a distinct nucleocytoplasmic shuttling during induced apoptosis. NPM was co-localized with Bax, Bcl-2 and their colocalization regions changed during the process of apoptosis. Conclusion NPM is a nuclear matrix associated protein. Its altered expression and localization, as well as its co-localization with apoptosis regulating proteins suggest the important regulative functions in apoptosis of EC9706 cells.