人肺巨细胞癌蛋白质组的二维电泳和计算机图象分析

为优化用于蛋白质组研究的二维电泳技术和计算机图象分析技术 ,以及初步分析比较与肿瘤细胞转移相关的蛋白质 ,以人肺巨细胞癌 (PLA- 80 1 - D、C)高、低转移株作为研究对象 ,应用 IPG-phor进行第一向等电聚焦 ,随后 ,在 Protein IPG conversion Kit上进行垂直 SDS- PAGE的分离 .利用光密度仪对银染的凝胶扫描 ,通过 PDQuest软件进行蛋白斑点检测和配比 .结果表明 :(1 )应用 IPGphor,采用样品直接加入重泡胀溶液的形式 ,增大了溶解性 ,缩短聚焦时间、增大样品负荷量 (分析型 ) ,提高了分辨率 .(2 )比较宽 (p H=3～ 1 0 L)、窄 (p H=4～ 7L)范围 IPG胶条 ,窄 p H范围的 IPG胶条具有较高的分辨率 .(3)比较 PLA- 80 1 - C、D细胞蛋白图谱之间的差异 ,其相关系数为 0 .7339± 0 .0 2 91 ;仅在 PLA- 80 1 - C株出现的蛋白为 1 79个 .

Proteome Research of Human Lung Giant Cell Carcinoma by Two-dimensional Gel Electrophoresis and Computer Image Analysis

To optimize the two|dimensional gel electrophoresis(2|DE)and computer assisted image analysis for the proteome research,and to compare the proteomic difference associated with tumor metastasis,the two cell sublines C,D of PLA|801,derived from a human lung giant cell carcinoma,were used as models.The rehydrated IPG strips containing protein samples were focuserphoresis chamber.After IEF,the equilibrated strips were transferred to the vertical SDS gel.The silver stained gels were scanned by the densitometer GS|710,which generated the digitalized images.The computer analysis including automatic detection and quantification of protein spots,as well as alignment,matching between the different 2|DE gels,was carried out by PDQuest software.The results demonstrated that:(1)In|gel sample application on the IPGphor electrophoresis chamber results in more sample loading and higher resolution.(2)The IPG strips with narrow pH range got better resolution than those with wide pH range do.(3)Image analysis made it possible to globally discriminate the difference of protein profile between two sublines PLA|801|C and D.