慢性粒细胞白血病不同病期蛋白质组分析优化策略

Better Strategy for Proteomic Analysis at Different Stages in Chronic Myeloid Leukemia

目的比较不同分组方法处理慢性粒细胞白血病（chronic myeloid leukemia，CML）急变前后骨髓细胞蛋白的差异表达，筛选急变相关蛋白。方法应用双向凝胶电泳（two—dimensional electrophoresis，2-DE）分析技术，比较急变前后三种分组方法（混合标本组、随机标本组、个体标本组）时骨髓细胞蛋白质的差异表达，并进行质谱鉴定。结果急变前后蛋白图谱匹配率在混合标本组为70．0678％；随机标本组两两配对比较，匹配率在43．2604％～58．3477％之间；个体标本组为74．0988％。混合标本组和随机标本组的相同差异蛋白点为103个，采用X^2检验P〉0．05。从三组共同表达的差异蛋白中选取了20个进行质谱分析，明确鉴定出15个蛋白质。结论蛋白质组学分析中，采用骨髓细胞混合标本分组策略可获得相关差异蛋白，且可有效消除个体差异，简化实验流程。

Objective To compare differentially expressed proteins in bone marrow cells at chronic and blast phases of chronic myeloid leukemia by means of different grouping methods, and to screen the proteins involving in the transformation. Methods Two- dimensional electrophoresis （2 - DE） was used to separate proteins and compare the changes of proteins expression in bone marrow cells at these two stages with three grouping methods, including the mixing samples group, the random samples group and the individual group. The differentially expressed protein spots were identified by peptide mass fingerprint in combination with database searching. Results The protein spots of the two stages were resolved with a match rate of 70.0678 % in the mixing samples group; 6 gel pages from CP and BC respectively were matched by pairs in the random samples group, and the range of match rate was from 43. 2604 % to 58.3477 %. The match rate was 74. 0988 % in the individual group. The common differentially expressed protein spots from the mixing samples group and the random samples group were 103, and X^2 test result was P〉0.05. Twenty protein spots were selected from the co - expressed different spots among these 3 groups for identification by peptide mass fingerprint in combination with database searching, and 15 spots were identified. Conclusion Distinct protein profiles can be obtained by using the mixing bone marrow cells in the comparative proteomics analysis. The distinction of individuals can be eliminated efficiently by this method and the steps of experiment can be simplified.