应用MALDI-TOF-MS技术分析转移和非转移黑色素瘤患者外周血清蛋白的差异表达

Differential expressions of peripheral serum protein in patients with metastatic and non-metastatic melanoma analyzed by MALDI-TOF-MS

摘要:

目的：应用蛋白质组学技术分析转移性和非转移性黑色素瘤患者外周血清中差异表达的蛋白质，初步探讨其对转移性黑色素瘤患者的诊断及治疗指导意义。方法：共收集101例患者外周血清，包括25例转移性黑色素瘤患者及76例对照者（31例非转移性患者和45例健康者）。从中随机抽取73例（18例转移患者，20例非转移患者和35例健康者）用于建立区分模型，其余28例（7例转移患者，11例非转移患者和10例健康者）用来对区分模型进行盲法验证。运用基质辅助激光解离飞行时间质谱技术，对外周血清蛋白质进行检测，应用Ciphergen蛋白质芯片软件对蛋白质谱进行分析。结果：黑色素瘤转移患者外周血清与对照组外周血清蛋白质谱相比，质荷比为2 598.33、3 646.79、4 605.48、8 603.10、7 756.64 m/z和9 598.82 m/z，共6个蛋白质峰呈表达有明显差异（P均=0.000），以此构建出转移性黑色素瘤的诊断模型，它的诊断敏感性为83.33%，特异性为74.55%。另外对该模型进行了盲法验证，其诊断转移性黑色素瘤的敏感性为71.43%，特异性为76.19%。结论：黑色素瘤转移患者外周血清蛋白质表达谱与对照组相比有明显变化，相对分子质量为2 598.33、3 646.79、4 605.48、8 603.10、7 756.64 m/z和9 598.82 m/z的蛋白质点可以作为黑色素瘤转移患者诊断的潜在生物学标志物。

Abstract:

Objective：To analyze the differential expressions of peripheral serum protein in patients with metastatic and non-metastatic melanoma by proteomic technology and to explore its clinical significance in the treatment of patients with metastatic melanoma. Methods：Totally 101 patients were selected and divided into metastatic melanoma group（n=25） and control group（n=76，31 cases of non-metastatic melanoma and 45 healthy cases）. Seventy-three cases（18 cases of metastatic melanoma，20 cases of non-metastatic melanoma and 35 healthy cases） were randomly extracted as diagnostic model group，the rest 28 cases（7 cases of metastatic melanoma，11 cases of non-metastatic melanoma and 10 healthy cases） were used to verify for model group by blind tests. Matrix assisted laser desorption ionization time of flight mass spectrometry（MALDI-TOF-MS） was applied to test peripheral serum protein. Ciphergen pro－tein chips were used to analyze protein profiling. Results：Mass-to-charge ratios of peripheral serum of patients with metastatic mela-noma against that of control group were 2 598.33，3 646.79，4 605.48，8 603.10，7 756.64 m/z and 9 598.82 m/z. Differences in expres－sions of six protein peaks were statistically significant（all P=0.000）. Diagnostic model of metastatic melanoma was built according to the statics above. Sensitivity of diagnostic model group was 83.33% and specificity was 74.55%. In addition，validation groups were used for this model by blind tests. Sensitivity of metastatic melanoma was 71.43% and specificity was 74.55% based on the model. Conclusions：There are significant changes in expression profile of peripheral serum protein between patients with metastatic melanoma and controls；proteins with relative molecular mass of 1 528.33，2 046.79，3 685.48，4 392.10，5 756.64 m/z and 11 670.82 m/z can be used as biological markers in the treatment of early metastatic melanoma.