骨肉瘤组织与良性骨肿瘤的比较蛋白质组学分析

Comparative proteomics analysis of human osteosarcomas and benign tumors of bone

目的:分析骨肉瘤与骨良性肿瘤差异蛋白质质点,寻找影响骨肉瘤恶性生物学行为的分子指标。方法:固相pH梯度双向凝胶电泳分离5例骨肉瘤组织与5例良性骨肿瘤组织总蛋白。Deep purple荧光染色,Im-ageMaster 2DE软件分析,对差异蛋白质点采用基质辅助激光解吸电离飞行时间质谱测定肽质指纹图谱并查询SWISS-PROT数据库。结果:获得重复性较好的双向电泳荧光染色图谱。图像分析检测骨肉瘤组织平均蛋白点数为1 386±101,良性骨肿瘤组织平均蛋白点数为1 270±202。初步鉴定出22个差异表达蛋白质点,其中15个在骨肉瘤中表达上调,以锌指蛋白133(zinc finger protein 133,Znf133)、laminB和T-复合多肽1(tailless complex poly-peptide 1,TCP-1)上调明显;7个在骨肉瘤中表达下调,其中蛋白激酶C抑制因子-1(protein kinase C inhibitor pro-tein 1,KCIP-1)表达缺失。结论:骨肉瘤组织与良性骨肿瘤的蛋白质表达存在明显差异,其中Znf133、laminB、TCP1、KCIP-1等可能在骨肉瘤的发生发展中发挥重要作用,有可能成为诊断骨肉瘤的分子标记物或治疗靶点。

AIM： To analyze the proteomie components of the tissue from human osteosareoma and benign tumor of bone, and to search the diagnostic markers of osteosareoma. METHODS ： Five samples of osteosareoma and 5 additive samples from benign bone tumor were analyzed by a series of methods ineluding immobilized pH gradient two - di- mensional polyaerylamide gel eleetrophoresis （2DE）, deep purple staining. The ImageMaster 2DE analysis -software, as well as peptide mass fingerprint based on matrix - assisted laser desorption/ionization time of flying mass speetrometry （MALDI- TOF- MS） and SWISS -PROT database searching were used for data processing. RESULTS： The average spots in osteosareoma and the benign tumor of bone were 1 386 ± 101 and 1 270 ±202, respectively. 22 peptide mass fingerprint （PMF） maps were obtained by MOLDI - TOF - MS and identified by searching the SWISS - PROT database. Compared with those in benign tumor of bone, 15 proteins were significantly up -regulated, especially zinc finger protein 133 （Znf133）, lamin B and tailless complex polypeptide 1 （TCP- 1 ）. 7 down -regulated proteins were observed, with the absence of protein kinase C inhibitor protein 1 （ KCIP - 1 ） in osteosarcoma. CONCLUSION ： The results suggest that an obviously differential proteomic expression exists between the osteosarcoma and benign tumor of bone. The overexpression of Znf133, lamin B, TCP1 and low - expression of KCIP - 1 may play an important role in the development of osteosarcoma, and may serve as diagnostic markers/therapeutic targets of osteosarcoma.