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在鉴定视黄酸(retinoic acid,RA)诱导人神经母细胞瘤SK-N-SH细胞分化的基础上,应用免疫细胞化学、选择性抽提和蛋白质组学分析技术,对SK-N-SH细胞诱导分化过程中核基质蛋白组成变化进行了系统研究.实验结果显示,经1μmol/L RA处理后SK-N-SH细胞呈极性状,伸出较长的轴突样突起,胞体逐渐变小变圆.免疫细胞化学结果显示,处理后神经细胞特异表达的蛋白synaptophysin、NSE、MAP2的表达量都较对照组有明显增强.双向凝胶电泳分析显示,在RA诱导SK-N-SH细胞分化前后存在52个差异表达的核基质蛋白,经质谱分析,鉴定了其中的41个蛋白.蛋白印迹杂交进一步确证了诱导分化差异表达核基质蛋白中nucleophosmin和prohibitin等的表达变化.研究结果表明,1μmol/LRA对SK-N-SH细胞具有显著的诱导分化作用,在SK-N-SH细胞分化过程中,其核基质蛋白组成发生了明显变化.这些变化对于揭示人神经母细胞瘤细胞癌变与逆转机制和肿瘤细胞增殖与分化调控机理均有十分重要的意义,从而为研究神经系统正常发育过程及神经系统疾病的发病机理提供科学依据.
On the basis of identifying the differentiation of human neuroblastoma SK-N-SH cells induced by retinoic acid (RA), immunocytochemistry, selective extraction and proteomic analysis were used to detect the expression of SK-N-SH The results showed that SK-N-SH cells exhibited a polar morphology with prolonged axon-like projections after 1 μmol / L RA treatment, and the cell bodies gradually became smaller Immunocytochemistry showed that the expression of synaptophysin, NSE and MAP2, which were specifically expressed in neural cells, were significantly increased after treatment compared with the control group.Analysis by two-dimensional gel electrophoresis showed that before and after the differentiation of SK-N-SH cells induced by RA Fifty-two differentially expressed nuclear matrix proteins were identified and 41 of them were identified by mass spectrometry.Western blotting confirmed the expression changes of nucleophosmin and prohibitin in differentially differentiated nuclear matrix proteins.The results showed that 1 μmol / LRA has a significant induction and differentiation effect on SK-N-SH cells, and its nuclear matrix protein composition changes significantly during the differentiation of SK-N-SH cells.These changes are of great significance for revealing the expression of human neuroblastoma Cell carcinogenesis and reversal mechanisms and tumor cell proliferation and differentiation regulation mechanisms are of great significance, thus providing a scientific basis for the study of the normal development of the nervous system and the pathogenesis of nervous system diseases.