The Key Laboratory of Ministry of Education for Cell Biology and Tumor Cell Engineering, School of Life Science, Xiamen University, Xiamen 361005, PR China.
J Cell Biochem. 2010 Sep 1;111(1):67-74. doi: 10.1002/jcb.22663.
In this article, we selectively extracted the nuclear matrix and intermediate filament system of human neuroblastoma SK-N-SH cells pre- and post-treated with retinoic acid (RA). The distribution of nucleophosmin (NPM) in the nuclear matrix and its colocalization with several products of related genes were investigated. Results from two-dimensional gel electrophoresis and MALDI-TOF showed that NPM was a component of the nuclear matrix and its expression in SK-N-SH cells post-treated with RA was down-regulated. Immunofluorescent microscopy observations further showed that NPM was localized in the nuclear matrix of SK-N-SH cells, and its expression level and distribution were altered after treatment with RA. The colocalization of NPM with c-myc, c-fos, p53, and Rb in SK-N-SH cells was observed under a laser scanning confocal microscope, but the colocalization region was changed by RA. Our results prove that NPM is a nuclear matrix protein, which is localized in nuclear matrix fibers. The colocalization of NPM with its related genes and oncogenes affect the differentiation of SK-N-SH cells. The expression of NPM and its distribution in the process of cell differentiation deserve more intensive investigation.
本文选择性提取了维甲酸(RA)预处理和后处理的人神经母细胞瘤 SK-N-SH 细胞的核基质和中间纤维系统。研究了核磷蛋白(NPM)在核基质中的分布及其与相关基因产物的共定位。二维凝胶电泳和 MALDI-TOF 的结果表明,NPM 是核基质的组成部分,RA 处理后的 SK-N-SH 细胞中 NPM 的表达下调。免疫荧光显微镜观察进一步表明,NPM 定位于 SK-N-SH 细胞的核基质中,RA 处理后其表达水平和分布发生改变。在激光共聚焦显微镜下观察到 NPM 与 c-myc、c-fos、p53 和 Rb 在 SK-N-SH 细胞中的共定位,但 RA 改变了共定位区域。我们的结果证明 NPM 是一种核基质蛋白,定位于核基质纤维中。NPM 与其相关基因和癌基因的共定位影响 SK-N-SH 细胞的分化。NPM 的表达及其在细胞分化过程中的分布值得更深入的研究。