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目的应用飞行时间质谱技术结合蛋白芯片筛选卵巢恶性肿瘤患者的血清标志物。方法选择广西医科大学附属肿瘤医院自2003年1月—2005年5月间收治的卵巢恶性肿瘤患者99例(恶性肿瘤组),卵巢良性肿瘤患者21例(良性肿瘤组);另取体检中心健康妇女87例(正常组)作为对照。分别采用IMAC3型固定金属亲和芯片和WCX2型弱阳离子交换芯片检测各组血清标本,应用Ciphergen蛋白芯片3．2．0软件采集3组标本的蛋白峰进行比较,蛋白峰强度相差0．5倍以上者定义为差异蛋白。同时应用Biomarker Pattern 5．0软件建立由IMAC3和WCX2型芯片筛选结果共同组成的诊断模型,扩大样本量,对该诊断模型进行盲法验证。应用本诊断模型筛选出潜在的卵巢恶性肿瘤血清标志物。结果将恶性肿瘤组(随机选取其中41例)与正常组(随机选取其中39例)血清标本比较,其中31个蛋白为差异蛋白(P<0．05)。建立诊断模型后,扩大样本量,分别取恶性肿瘤组中另外58例卵巢恶性肿瘤患者的术前血清和正常组中另外48例健康妇女的血清进行盲法验证,结果显示,其敏感度为100％,特异度为98％。将恶性肿瘤组与良性肿瘤组血清标本比较,发现质荷比为7769的蛋白为卵巢恶性肿瘤的潜在标志物。结论应用飞行时间质谱技术结合固定金属亲和芯片和弱阳离子交换芯片,可有效筛选血清中特异性蛋白标志物,为建立卵巢恶性肿瘤的诊断模型提供可靠的技术平台。
Objective To screen serum markers of ovarian cancer patients by using time-of-flight mass spectrometry combined with protein microarray. Methods 99 cases of ovarian cancer patients (malignant tumor group) and 21 cases of benign ovarian tumor (benign tumor group) were enrolled in the Affiliated Tumor Hospital of Guangxi Medical University from January 2003 to May 2005. Another physical examination center 87 women (normal group) as a control. The serum samples of each group were detected by IMAC3 type immobilized metal affinity chip and WCX2 type weak cation exchange chip. The protein peaks of three groups were compared by using Ciphergen protein chip 3.2.0 software, the peak intensity of protein was different by 0.5 times The above is defined as the differential protein. At the same time, Biomarker Pattern 5.0 software was used to establish a diagnostic model consisting of IMAC3 and WCX2 chip screening results. The sample size was expanded and the diagnostic model was validated blindly. Use this diagnostic model to screen out potential ovarian cancer serum markers. Results The malignant tumor group (41 randomly selected) and the normal group (39 were randomly selected) serum samples, of which 31 proteins were differential proteins (P <0.05). After the establishment of the diagnostic model, the sample size was expanded, and the sera of another 58 patients with malignant ovarian tumor in preoperative serum of the malignant tumor group and 48 healthy women in the normal group were blind-tested. The results showed that the sensitivity was 100 %, Specificity of 98%. The malignant tumor group and benign tumor serum samples compared and found that the mass-to-protein ratio of 7769 for ovarian cancer is a potential biomarker. Conclusion Time-of-flight mass spectrometry combined with immobilized metal affinity chip and weak cation exchange chip can effectively screen specific protein markers in serum and provide a reliable technical platform for the establishment of a diagnostic model of ovarian cancer.