骨肉瘤与正常骨组织的蛋白质组学分析

目的 比较骨肉瘤与正常骨组织蛋白质组学的差异,探寻骨肉瘤特异性标志物.方法 分别提取5例骨肉瘤肿瘤标本和5例正常骨组织总蛋白,进行双向电泳,重复3次；银染、扫描后进行ImageMaster 2-DE Platinum 6.0软件分析以发现差异蛋白并对差异蛋白质点进行MALDI-TOE TOF质谱鉴定,获取肽质指纹图谱后应用NCBI数据库查找匹配蛋白质.结果 获得重复性较好的双向电泳银染图谱,骨肉瘤组织蛋白质点数约为(1 296±179),正常骨组织蛋白质点数约为(1108±162),明显差异的蛋白质点25个,其中10个被成功鉴定,6个蛋白质在骨肉瘤中表达上调,以谷胱甘肽转移酶升高最为突出;4个蛋白质在骨肉瘤中表达下降,以核苷二磷酸激酶下降最明显.结论 蛋白质组学的分析能成功发现骨肉瘤与正常骨组织之间的差异蛋白质,为寻找骨肉瘤特异标志物提供依据.

Research for Proteomics Discrepancies between Osteosarcoma and Normal Bone Tissue

Objective
To compare the proteomics discrepancies between osteosarcomas and normal tissue of bone for discovering specific
markers for osteosarcoma.MethodsTotal protein was extracted from five samples of osteosarcoma and five samples of
normal bone,respectively.Then dielectrophoresis was conducted three times,repeatedly.The differential proteins were
found out by ImageMaster 2-DE software analysis after silver staining and scanning.Then,
MALDI-TOF/TOF mass spectrometric identification of differential protein spots was conducted.Get access to the
peptide mass fingerprints and query to find out matching proteins in the NCBI database.ResultsDielectrophoresis
silver staining map had good repeatability.Protein spots of osteosarcomas and normal bone were approximately(1 296±
179),(1 108±162),respectively,and 25 significant different proteins were discovered,of which 10 spots were
identified successfully.Six proteins were significantly up-regulated in osteosarcom,especially the level of
glutathiones transferases（GST）.Four down - regulated proteins were observed，especially the level of nucleoside
diphosphate kinase（NDPK）.ConclusionThe different proteins between osteosarcoma and normal tissue can be found by
proteomics analysis which provide the basis for searching the specific marker for osteosarcoma.