小儿神经母细胞瘤蛋白质组的初步探索

A preliminary exploration for the proteome of children neuroblastoma

目的应用蛋白质组学方法筛选与小儿神经母细胞瘤（NB）相关的差异表达蛋白质。方法收集2011年7～12月解放军总医院小儿外科经穿刺活检病理证实为Ⅳ期NB的标本3例及其化疗后术后切除肿瘤标本3例，患儿平均年龄3．17岁。对化疗前后NB组织全蛋白进行二维凝胶电泳，应用超高效纳升液相色谱一电喷雾串联质谱进行蛋白质斑点鉴定和生物信息学分析。结果成功建立了组织化疗前后双向电泳图谱，应用图像分析软件发现化疗后有7个差异表达明显的蛋白质斑点，其中有2个蛋白质表达上调，分别为Nm23蛋白和神经多肽h3；5个蛋白质表达下调，分别为癌蛋白18、热休克蛋白27、线粒体烯酰脂辅酶A、过氧化物酶1和过氧化物酶3。结论应用二维凝胶电泳和超高效纳升液相色谱一电喷雾串联质谱能鉴定小儿NB化疗前后的差异表达蛋白。

Objective To screen the differential expression proteins in the children with neuroblastoma （NB） by proteomics tools. Methods Thi＇ee specimens were collected from the patients diagnosed IV stage NB by biopsy at Department of Pediatric Surgery of Chinese PLA General Hospital in Beijing from July to December ,2011. Another three specimens were acquired from the same patients underwent tumor excision. Average age was 3.17 years. Proteins in neuroblastoma before and after chemotherapy were separated by two dimensional gel electrophoresis, analyzed by high performance liquid chromatography-electrospray tandem MS （Nano-UPLC-ESI- MS/MS）. Results After two dimensional gel electrophoresis, we obtained the maps about tissues before and after chemotherapy. There were 7 differential protein spots identified by using the Image Master two dimensional gel electrophoresis software,in which 2 were up-regulated, including Nm23 protein and neuropolypeptide h3,5 were down-regulated after chemotherapy, including stathmiul, heat shock protein 27, mitochondrial short-chain enoyl-coenzyme A, peroxiredoxin 1 and peroxiredoxin 3. Conclusion The differential expression proteins of children neuroblastoma before and after chemotherapy were successfully identified by two dimensional gel elec- trophoresis and Nano-UPLC-ESI-MS/MS.