毛细管电泳-激光诱导荧光检测法分析胃癌组织及癌旁正常组织蛋白质的差异性

胃癌是临床常见的恶性肿瘤之一。近年来寻找肿瘤相关特异蛋白质是蛋白质组学研究的热点。本文通过考察毛细管动态涂层方法、筛分介质聚环氧乙烷(PEO)的浓度、缓冲液的pH值、分离电压、温度及荧光染料对分离效果的影响,建立了毛细管电泳-激光诱导荧光法分离胃癌组织及癌旁正常组织蛋白质的方法;通过分离检测,获得两者的蛋白质指纹图谱。经分析,两者的指纹图谱相似度达到0.8以上,差异蛋白质分子质量集中在50000~100000 Da之间,提示某些小分子蛋白质可能是和肿瘤发生相关的特异蛋白质,从而缩小了特异性分子标记物的筛选范围。病理组织学分型及蛋白质电泳峰数目的统计结果验证了该方法的可靠性。该方法具有临床应用的潜力。

Analysis of difference of proteins of gastric cancer tissue and adjacent normal tissue by capillary electrophoresis with laser-induced fluorescence detection

Gastric cancer is one of the familiar malignant tumors in clinical study. Research on tumor biomarkers has increased noticeably in recent years. An experimental method of protein separation by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) was established and improved. A series of effects on capillary electrophoresis (CE) were studied, such as the dynamic coating method of capillary, concentration of polyethylene oxide (PEO) as sieving medium, pH of running buffer, separation voltage, temperature and fluorescent dye. Then the optimized method was established for the determination of gastric cancer tissue and adjacent normal tissue. According to the analysis of the protein fingerprints, the results showed that the similarity of protein was up to 0.8. The molecular masses of differential proteins were almost between 50000 Da and 100000 Da. It indicated that these proteins might be potential biomarkers for cancer diagnosis which may reduce the searching scope. By statistical analysis of histological classification and electrophoresis peak numbers, this method was demonstrated reliably, and had the potential for clinical applications.