双向电泳分析结晶型NiS诱发人支气管上皮细胞恶变后的蛋白表达差异

Differential Protein Expression in Crystalline Nickel Sulfide-induced Malignant Transformed Cells by Two Dimensional Electrophoresis

[目的]建立双向凝胶电泳分析方法 ,比较结晶型NiS诱发人支气管上皮细胞恶变后的蛋白表达差异。[方法]采用一步法分别提取人支气管上皮细胞系16HBE细胞和结晶型NiS诱发该细胞系的恶性转化细胞N_2细胞的可溶性总蛋白 ,经双向凝胶电泳分离、银染显色、用双向电泳凝胶图像分析软件ImageMaster2D3.10分析两者之间可溶性蛋白的差异表达。[结果]获得了分辨率和重复性较好的双向电泳银染图谱。图像分析显示在恶性转化细胞中检测到19个新的蛋白点 ,同时有47个蛋白点消失 ,有140个蛋白点在两种细胞中表达量有显著差异,其中89个蛋白点在N_2细胞中表达升高 ,51个蛋白点在N_2细胞中表达降低。[结论]双向电泳技术分析NiS诱发细胞恶性转化后蛋白质组发生了变化 。

Objective To establish a method of the two-dimensional electrophoresis(2-DE)for investigating the differential protein expression in human bronchial epithelial malignant transformed cells induced by crystalline nickel sulfide.[Methods] The whole soluble proteins from16HBE and malignant transformed N-2cells were extracted by one step method and separated using2-DE.The2-DE gels were stained with silver staining.Different expressions of the whole soluble proteins were analyzed using the analysis software Imagemaster2-D3.10.[Results]The good2-DE pattern including resolution and reproducibility was obtained.The differential proteome expression analysis found that19protein spots were specifically detected,and47protein spots disappeared in N-2cells,compared with those in16HBE cells.In addition,the expression level of140protein spots was different among them,of which the expression level of89protein spots was up-regulated,51protein spots down-regulated in N-2cells.[Conclusion] The differenˉtial proteome expression in crystalline nickel sulfide-induced malignant transformed cells is different from that in16HBE cells using2-DE.It provides opportunity to identify proteins related to nickel carcinogenesis in further study.