运用基质辅助激光解吸电离飞行时间质谱技术探索建立鼻咽癌肝转移特异血清多肽谱诊断模型  

Establishment of a specific serum proteomic profile model for liver organ-specific metastasis of nasopharyngeal carcinoma by matrix-assisted laser desorption/ionization time of flight mass spectrometry

作　　者：潘长穿[1] 尹序德[1] 喻璟瑞[1] 张智慧[1] 吴沛宏[2] 夏云飞[3] 

机构地区：[1]四川省肿瘤医院/研究所,四川省第二人民医院肿瘤内科,四川成都610041 [2]中山大学肿瘤防治中心影像介入科,华南肿瘤学国家重点实验室,广东广州510060 [3]中山大学肿瘤防治中心放疗科,华南肿瘤学国家重点实验室,广东广州510060

出　　处：《肿瘤》2013年第9期814-819,共6页Tumor

基　　金：四川省卫生厅科研课题(编号:120029)

摘　　要：目的：探讨鼻咽癌肝转移患者与鼻咽癌其他器官转移、鼻咽癌不发生远处转移及其他恶性实体肿瘤肝转移患者的血清蛋白谱差异，建立特异性的鼻咽癌肝转移患者血清多肽谱诊断模型。方法：50例随访时间〉3年的鼻咽癌患者中，发生肝转移者16例作为研究组，肝外远处转移患者16例和不发生远处转移的患者18例作为对照组；同期纳入其他恶性实体肿瘤肝转移患者14例，也计入对照组。收集所有患者的血清标本，通过特异的液体磁珠分选血清后，进行基质辅助激光解吸电离飞行时间质谱（matrix-assisted laser desorption／ionization time of flight mass spectrometry，MALDI-TOF-MS）技术分析，得到血清多肽谱。采用专用的生物信息学分析软件ClinProtTools进行差异分析，并用工具包中的数据挖掘算法（神经网络算法、快速分类算法、支持向量机算法和遗传算法）及分类决策树来建立鼻咽癌肝转移的血清多肽谱诊断模型。结果：MALDI-TOF-MS法检测出鼻咽癌肝转移和鼻咽癌不发生远处转移的患者中差异蛋白峰有28个，鼻咽癌肝转移和鼻咽癌肝外转移患者的差异蛋白峰有9个，鼻咽癌肝转移和非鼻咽恶性实体肿瘤肝转移患者的差异蛋白峰有45个，鼻咽癌肝外转移和鼻咽癌不发生远处转移的患者中差异蛋白峰有10个。通过比较性蛋白质组学分析筛选出鼻咽癌肝脏特异性转移的4个血清蛋白质谱峰的质荷比分别为4155.34、4194.87、4210.78和4249.56，建立诊断鼻咽癌肝转移和鉴别诊断鼻咽癌肝外转移、鼻咽癌不发生远处转移以及非鼻咽恶性实体肿瘤肝转移的模型，各组模型的诊断和鉴别诊断的理论识别率均为100％，交叉验证的预测能力高达73.3％～100％。结论：鼻咽癌肝转移患者存在特异性蛋白表达，建立的特异性血清多肽诊断模型在鼻咽癌肝脏特异性转移诊断中有应用前景，为进�Objective： To investigate the specific serum peptide profile of LM （liver metastasis） associated with NPC （nasopharyngeal carcinoma） by comparing the patients who have NPC with LM and without LM and the patients with LM not from NPC, and to provide the model for diagnosis of LM from NPC. Methods： Pre-treatment serum samples from 50 patients who had pathologically confirmed NPC and 14 patients who had pathologically confirmed non-NPC with LM were collected and assayed by MALDI-TOF-MS （matrixassisted laser desorption/ionization time of flight mass spectrometry） analysis. During follow-up of morethan 3 years after radiotherapy, 16 NPC patients with LM （LM NPC group）, 16 NPC patients with non-LM （non-LM NPC group） and 18 NPC pateints without metastasis （non-M NPC group） were confirmed. Mass spectrographic data were analyzed with ClinProt software Tools. The specific serum peptide model of NPC-associated LM was established by using both data mining analysis and decision tree classification analysis. Results： Differential expressions of 28 peptide peaks were detected between LM NPC group and non-M NPC group, 9 peptide peaks between LM NPC group and non-LM NPC group, 45 peptide peaks between LM NPC group and LM non-NPC group, and 10 peptide peaks between non-LM NPC group and non-M NPC group. Using comparative proteomics analysis, 4 protein mass peaks （their mass to charge ratios were 4 155.34 m/z, 4 194.87 m/z, 4 210.78 m/z and 4 249.56 m/z, respectively）were identified as the liver-specific metastasis-associated protein peaks in NPC. The models based on the 4 sieved markers of NPC could discriminate LM NPC group from non-LM NPC group, non-M NPC group and non-NPC LM group. The recognition capability was 100.0% and the cross-validation of these models for differentiating the above 4 groups were 73.3%-100.0%. Condusion： NPC with LM has a specific serum peptide profile. The established specific serum peptide model may have certain application in the diagnosis of LM associated with NPC, a

关 键 词：鼻咽肿瘤 光谱法 质量 基质辅助激光解吸电离 肿瘤转移 肝 分子诊断技术 血蛋白质类 

分 类 号：R739.62[医药卫生—肿瘤] R739.63[医药卫生—临床医学]