应用三维鸟枪法串联质谱技术研究脊髓室管膜瘤及正常脊髓室管膜的蛋白表达谱

Study on proteomic profile of normal ependyma and ependymoma of spinal cord by using three-dia- mensional Shot-gun and MS/MS technology

目的探讨应用三维鸟枪法（Shot—gun）串联质谱分析技术研究脊髓室管膜瘤和正常室管膜蛋白表达谱的可行性。方法分别采集8例脊髓室管膜瘤标本和4例室管膜标本获取蛋白，进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS—PAGE），考马斯亮蓝染色、切胶和酶解，串联质谱获取肽质量指纹图谱并鉴定和生物信息学分析。结果室管膜组鉴定组2758蛋白，室管膜瘤组鉴定2721个蛋白。其中1631个蛋白质在肿瘤组和对照组均有表达，1090个蛋白质仅在肿瘤组中表达，1127个蛋白质仅在对照组中表达。结论SDS—PAGE三维Shot—gun串联质谱技术为定量比较蛋白组学研究脊髓室管膜瘤奠定了技术平台。

Objective To research fbcused on proteomic profile of normal ependyma and ependy- morea of spinal cord by using three-diamensional Stlot-gun and MS/MS technology. Methods Eight spinal ependymoma and four normal ependyma tissues were obtained through operations. The samples were pro- gressed total proteins extraction, sodium dodecyl sulfate polyacrylamide gel electropheresis （SDS-PAGE） electrophoresis, gels cutting, protein enzymolysis and MS/MS. The samples were progressed total proteins extraction. The identified proteins were analysis by bioinformatics. Results A normal spinal ependyma proteome consist of 2758 proteins while the ependymoma proteome consist of 2721. 1631 proteins were de- tected both in tumor group and in control group, 1090 proteins were only detected in tumor group while 1127 proteins only in control group. Conclusion All the proteomic profile studies by using three-diamen- sional SDS-PAGE and Shot-gun and MS/MS technology provide a foundation for the further tumor-genetical research of spinal ependymoma.