蛋白质组学技术在髓系白血病分化研究及白血病早期诊断中的应用

目的 探讨白血病细胞粒细胞系、单核细胞系两系分化的蛋白质组学研究,分析差异表达蛋白质在白血病筛查中的应用价值.方法 利用ATRA和甾体类新药(编号:NSC67657)诱导髓系白血病HL60细胞,构建白血病细胞粒细胞系和单核细胞系分化模型.采用双向电泳技术,分离HL60细胞向粒细胞系、单细胞系分化前后差异表达蛋白质分子,经MALDI-TOF MS鉴定.对EF1A1、TLE1、NME3这3个差异表达蛋白质分子进行RT-PCR和WB验证.收集临床5例白血病患者和1名健康人的骨髓标本,研究3个差异表达蛋白质分子在白血病患者骨髓细胞中的表达情况.结果 WB分析发现细胞分化前后NME3蛋白质表达下降(对照组A=0.227,NSC67657处理组A=0.079,ATRA处理组A=0.064,2种药物处理组与对照组比较差异有统计学意义,P均＜0.01).在4份白血病患者标本中NME3表达水平高于健康人骨髓标本(健康人A=0.082,2例慢性粒细胞白血病急变期患者A=0.274、0.269,急性单核细胞白血病患者A=0.297,1例慢性粒细胞性白血病患者A=0.258,与健康人比较差异有统计学意义,P均＜0.05).另1例慢性粒细胞白血病患者A=0.121,与健康人比较差异无统计学意义,P＞0.05.结论 蛋白质组学技术从基础研究到临床应用是个阶段性的过程,本研究通过对筛选所得分化相关蛋白质NME3在白血病患者骨髓中的表达分析,为后续蛋白质组学技术在白血病早期诊断中的应用奠定了基础.

The application of proteomic technology in differentiation and early diagnosis of myelocytic leukemia

Objective To figure out the differentially expressed proteins using proteome technology in leukemia cells induced into different lineages and investigate the application value in early screening of leukemia.Methods With induction of ATRA and NSC67657, the differentiation models was constructed using HL60 cells which has the potentiality to be induced into different lineages by different inducers.Then the differentially expressed proteins in the process of differentiation was separated using two-dimensional electrophoresis and identified using MALDI-TOF MS.The expression of 3 proteins FE1A1, TLE1, NME3 were chosen to be verified in myeloid samples of 5 leukemia patients and 1 normal volunteer using RT-PCR and WB.Results WB showed that NME3 was differentially expressed after both granulocytic and monocytic differentiation( Normal A value = 0.227, NSC67657 A value= 0.079, ATRA A value = 0.064, P ＜ 0.01 ).However, only in 4 of 5 tested patients' myeloid samples, the NME3 protein expression were differentially expressed compared to the normal myeloid sample( Normal A value = 0.082,2 acute leumia transferred from chronic granulocytic leumia A value = 0.274,0.269, acute monocytic leukemia A value = 0.297, one patient with chronic granulocytic leukemia A value = 0.258.There was significant difference between normal and leukemia group, P ＜0.05 ).A value was 0.121 for another patents with chronic granulocytic leukemia The NME3 protein expression was not differentially expressed compared to the normal myeloid sample,P ＞0.05.Conclusions It is still a long way to go for proteome technology from basic research to clinical application.However, the identification of NME3 protein related to differentiation in leukemia patients' myeloid samples had set the foundation for the early diagnosis of leukemia using proteome technology.