应用蛋白质组学筛选宫颈癌患者尿液中的肿瘤标志物

Application of Proteomics to Screen Urinary Biomarkers in Women with Cervical Cancer

通过比较健康女性和宫颈癌患者的尿蛋白质组,发现并分析差异表达蛋白,从中筛选潜在的宫颈癌的标志物。研究对象由43名宫颈癌患者(CC)和47名健康女性(HW)组成。用超速离心法沉淀尿蛋白,再用一维凝胶电泳(SDS-PAGE)与液相色谱-质谱联用技术(LC-MS/MS)鉴定尿液中的蛋白质,蛋白质定量采用无标定量。比较患者尿蛋白质组、健康对照的尿蛋白质组和宫颈癌组织蛋白质组,有1910个蛋白质是患者和健康对照共有的尿蛋白,这其中有746个蛋白质也存在于宫颈癌组织蛋白质组。在这746个蛋白质中找到84个上调蛋白和82下调蛋白。通过生物信息学分析发现牛皮癣素(S100A7)和癌胚抗原相关细胞黏附分子8(CEACAM8)是宫颈癌尿液样本独有蛋白质。在验证组的70例样本中,双盲法测试S100A7、CEACAM8以及两者联合诊断宫颈癌的敏感性能达到73%、87%、93%。结果提示,宫颈癌患者的尿蛋白质组与健康女性的尿蛋白质组不同,并且S100A7和CEACAM8可以作为宫颈癌潜在的肿瘤标志物。

To explore the differential proteome pattern and screen biomarkers in urine samples from cervical cancer patients,urine samples from 43 patients suffering from cervical cancer（CC） and 47 healthy women（ HW） were collected for further proteomic analysis. Urine proteins precipitated by ultracentrifugation were subjected to SDS-PAGE separation and LC-MS / MS（ liquid chromatograph-mass spectrometer / mass spectrometer）analysis by using the label-free protein quantification assay. By comparing the urine proteomes between CC,HW and cervical cancer tissue groups,over 1910 proteins were mutually exclusive in CC and HW groups,while 746 proteins were found in all three groups. In the subset of 746 proteins,84 up-regulated proteins and 82 downregulated proteins in CC group when compared with the HW group. Further bioinformatics analysis showed that S100A7（ S100 calcium binding protein A7） and CEACAM8（ carcinoembryonic antigen-related cell adhesion molecule 8） were specific to CC. For further validation,LC-MS / MS was applied quantification on 70 samples.The combined analysis of S100A7 and CEACAM8（ S100A7 ＋ CEACAM8） can improve the sensitivity of cervical cancer diagnosis from 73%（ S100A7） and 87%（ CEACAM8） to 93%. The urine proteome of cervical cancer was different from urine proteome of healthy women be envied. S100A7 and CEACAM8 were potential biomarkers for cervical cancer.