应用液体蛋白芯片-飞行时间质谱技术诊断乳腺癌的临床价值

Clinical Value of Diagnosing Breast Cancer in Serum Using Magnetic Beads and MALDI-TOF-MS

目的应用弱阳离子磁珠-飞行时间质谱-Clin Prot系统联合筛选乳腺肿瘤患者与健康对照女性血清中的差异蛋白,建立乳腺癌诊断模型,并初步探究该模型诊断乳腺肿瘤的临床应用价值。方法采用弱阳离子交换磁珠吸附提取血清标本(乳腺癌120例,健康对照40例)中的表达蛋白质/多肽,应用基质辅助激光解析串联飞行时间质谱仪捕获血清蛋白谱图,利用Clin Prot v3.0软件分析质谱图信息,找到2组间具有明显差异蛋白质/多肽,建立乳腺癌诊断模型,并选择乳腺癌和健康对照标本各20例进行双盲验证。同时检测纳入标本血清中癌胚抗原(CEA)和癌抗原153(CA153)的含量,探究该模型对乳腺癌血清学诊断的临床价值。结果比对病例组和对照组的蛋白质谱图,得到35个具有明显差异的蛋白峰(P<0.05),其中7个在乳腺癌中表达上调,28个表达下调。根据遗传算法选取Mass 1 569、3 934、5 750、7 765建立乳腺癌诊断模型,该模型诊断乳腺癌和健康女性的灵敏性为95.0%,特异性为92.5%。双盲验证结果显示灵敏性为85.0%,特异性为90%。该模型对乳腺癌的诊断效能高于CEA和CA153的联合诊断。结论应用液体芯片-飞行时间质谱技术-Clin Prot系统能够检测乳腺癌血清中的差异蛋白,同时能够建立灵敏性、特异性均较好的乳腺癌诊断模型,为乳腺癌诊断开拓了新的途径。

Objective To use liquid protein chip matrix-assisted laser desorption/ionization time-of-flight mass spectrometry（MALDITOF-MS） of the ClinProt system to establish a diagnostic model for breast cancer and to explore its clinical diagnostic value. Methods Serum samples from 120 patients with breast cancer and 40 healthy controls were collected at the Department of Thyroid and Breast Surgery in the General Hospital of the Jinan Military Command Region. The weak cation exchange（WCX） beads combined with the MALDI-TOF-MS technique were used to detect differentially expressed proteins between breast cancer patients and controls. ClinProt v3.0 software was utilized to establish a diagnostic model for breast cancer. Further evaluation of the validation group in a blind test was used for recognition accuracy. Results Thirty-five differentially expressed proteins in serum were identified by comparing serum proteomic spectra between breast cancer patients and healthy individuals,including seven upregulated proteins and 28 downregulated ones. Four differentially expressed proteins were selected to build a diagnostic model of breast cancer. The sensitivity and specificity were 95.0% and 92.5%,respectively. Then,20 serum samples from patients with breast cancer and 20 healthy control samples were verified by the model. The recognition accuracy of the breast cancer group was 94.4%. Conclusion Use of the WCX beads-MALDI-TO-MS-ClinProt system could enable identification of novel biomarkers and establish a diagnostic model for breast cancer. It provides a new diagnostic method for breast cancer.