摘要
目的:对传统的双向凝胶电泳技术和计算机图像分析技术进行优化并将其应用于胃癌蛋白质组研究,提高实验数据分辨率和实验结果的可重复性。方法:提取胃癌组织中蛋白质,对以固相pH梯度为第一向双向电泳的关键因素与环节(如样品处理、电泳参数和凝胶浓度等)进行一系列优化。进一步采用双向凝胶电泳分离胃癌组和正常组总蛋白质,银染显色,并通过优化的计算机图像分析软件分析电泳图谱。结果:重复性实验结果显示同组样品在3次不同实验中所得蛋白质斑点数目相对标准差(变异系数平均值%):胃癌组和正常组分别为(23.00±10.11)、(20.33±9.90)。变异系数范围(%):胃癌组和正常组分别为3.80～60.10、2.70～56.89。同一蛋白质斑点在3次实验中等电点、分子量的相对标准差分别为(8.76±5.14)%,(13.00±4.22)%和(10.84±9.16)%。获得了优于传统实验方法的分辨率和可重复性的双向凝胶电泳图谱。结论:优化的双向凝胶电泳和图像分析技术适合于胃癌蛋白质组研究。
Objection： To establish and optimize the two-dimensional gel electrophoreses （2- DE） and the computer image analysis technique for the proteome research of the stomach cancer, To increase its resolution and its repetition in order to gain a kind of new method for analyzing stomach cancer proteome. Methods： The proteins of stomach cancer were extracted. Immobilized pH gradient was taken as the key factor and link in the isoelectric point focus. For instance, sample treatment, electrophoresis parameter and gel concentration were got on with a series of optimization. 2-DE was further adopted to separate the total protein of normal term and of stomach cancer term. Then the 2- DE chart was analyzed by silver and image analysis software. Results： The result of repetitive experiments showed that in three different experiments the amount of protein spots of the same term sample deviated from the relative standard as following （the average of variation coefficient %）. Stomach cancer and the normal term was （23.0±10.11）and （20.33±9.90） respectively. The range of variation coefficient （%）：Stomach cancer and the normal term were 3.80-60.10 and 2.70-56.89 respectively. The isoelectric point and molecular weigh of the same protein spots in three times experiments deviates from relative standard as following：（8.76±5.14） %, （13.00±4.22） % and （10.84±9.16） % respectively. Therefore, better resolution and repetitive 2-DE chart were obtained. Conclusion： The establishment and the optimization of proteome 2-DE are qualified to the expression study of stomach cancer proteome differentials.
出处
《海南医学院学报》
CAS
2005年第5期381-385,共5页
Journal of Hainan Medical University
基金
海南医学院苗圃基金资助课题(NO.2004117)
关键词
胃肿瘤
癌
蛋白质组
电泳
凝胶
双向
人
Gastric tumor
cancer
proteome
electrophoreses, gel, two -dimensional
person
