摘要
目的:通过分析卵巢癌患者血清中的蛋白质组分,寻找可能的肿瘤相关蛋白并对该蛋白鉴定,以期确定其作为卵巢癌血清学诊断的特异标志物。方法:采用双向凝胶电泳方法分离6例卵巢癌患者及2例正常健康妇女血清总蛋白,经考马斯亮蓝染色后对比找出差异蛋白斑点,该蛋白斑点经蛋白酶水解后进行液质联用串联质谱(LC-MS/MS)分析,用SEQUEST搜索软件查询Finngan公司提供的FASTA格式蛋白序列数据库。采用WesternBlotting、ELISA、斑点印迹及亲和层析等方法对目标蛋白作进一步的分析。结果:经双向电泳分离,卵巢癌患者血清中有3-4个组分显示含量高于正常健康妇女血清,选其中最明显的一个差异斑点进行质谱分析,对库检索确定为结合珠蛋白。进一步的分析证实,卵巢癌患者血清中结合珠蛋白总含量有高于正常健康妇女的趋势。此外,通过凝集素斑点印迹法检测发现,岩藻糖基化结合珠蛋白成分明显增加是该蛋白总量增高的主要特性。结论:检测血清结合珠蛋白总量以及特异的岩藻糖基化组分可能成为卵巢癌的血清诊断指标之一,可与其它检测方法相结合,提高卵巢癌早期诊断的有效性。
Objective: To find potential serum biomarkers by analyzing the protein omiccomponents of the sera from ovarian cancer patients and characterizing the potential tumor associated proteins in order to establish the protein as the specific serum marker for ovarian cancer diagnosis. Methods: Two - dimensional polyacryl-amide gel electrophor - esis (2DE) were used to separate the sera from the patients with ovarian cancer. After Coomassie Brilliant Blue staining, the target proteins were analysed by LC - MS/MS and the protein sequences were searched in database. The candidate biomarker was further analysed by Western Blotting, ELISA, Dot - blotting and affinity chromatograpy. Results: Several proteinomic components were found significantly increased in patient's serum. The most striking one was identified as haptoglobin (Hp) by LC -MS/MS. Further identification by Western Blotting and ELISA indicated that Hp was higher in cancer serum than that in normal. The significant increase of fucosylated haptoglobin in cancer serum was also observed. The method of lectin affinity Dot - blotting was established to measure fucosylated haptoglobin in serum. Conclusions: Haptoglobin and fucosylated haptoglobin are potential serum biomarkers. They are useful for o-varian cancer diagnosis when combined with other test markers
出处
《中国计划生育学杂志》
北大核心
2005年第4期230-233,共4页
Chinese Journal of Family Planning
