Proteomics Research Of CD34~+Cells From Patients With Acute Myeloid Leukemia Of M2Subtype

BACKGROUND and AIMAcute myeloid leukemia (AML) is a hematopoietic malignant disease which originates from a small group of abnormal cells called leukemia stem cells(LSCs).LSCs have the ability of self-renew and differentiation,they are suggested to be the cause of leukemogenesis and recurrence of AML.However,little is presently known about the characteristics of the LSCs in detail.Proteomics is defined as a techniques which takes protein of life functions unit as a breakthrough point and directly studies the composition and regulation of a protein.As a simple,quick and high throughout technology,proteomics plays an important role in the pathogenesis of research,diagnosis and therapy of tumor.Therefore,in this study, CD34+cells from the patients with AML-M2subtype leukemia were chose as a research object to looking for specificity and or associated protein molecules in order to explore the molecular mechanisms and the targets for molecular diagnosis and therapy of the AML-M2subtype leukemia.METHODSFirst of all, CD34+cells were positively sorted from the both patients with AML-M2subtype leukemia at diagnosis and control subjects of the mobilized peripheral blood of the donors of hematopoietic stem cell transplant by the method of magnetic activated cell sorting (MACS) followed by purity analysis of the sorted CD34+cells by flow cytometry.Then,proteins of the sorted CD34+cells were separated by fluorescence two-dimensional difference in-gel electrophoresis (2-D DIGE),the gel map was obtained with the Typhoon scanner,and the potent protein spots of differential expression were identified by means of the image analysis of DeCyder software. Finally, he candidate differentially expressed proteins spots were analyzed by mass spectrometry and bioinformatics.RESULTSAfter MACS sorting,the average purity of CD34+cells from the patients with AML-M2subtype leukemia was98%,while the average purity of CD34+cells from the control subjects was92%.The result of2-D DIGE analysis indicated that twenty-five potent proteins spots showed differential expression with one point one fold at least difference between the patients with AML-M2subtype leukemia and the control subject.Twenty-five potent proteins spots were further studied by mass spectrometry,and fifteen proteins peptide mass fingerprinting were obtained.Bioinformatics result exhibited that these proteins spots may represent heat shock protein (HSP)90beta,type2HSP90alpha,HSP70analogues,HSP70-1,HSP70-2,transferring,serotransferrin precursor, proliferating cell nuclear antigen,the calcium binding proteins,calreticulin precursor,pyruvic acid,pyruvate kinase M2,aldehydes reductase, nicotinamide adenine dinucleotide phosphate+,protein of PRO2619and other unknown proteins.these proteins are either associated with cellular and energy metabolism,or apoptosis,drug resistant,signal transduction, cell proliferation.CONCLUSIONBy MACS techniques,proteomics and bioinformatics analysis, in this study differentially expressed proteins have been found in the CD34+cells from the patients with AML-M2subtype leukemia,which may be associated with self-renew,differentiation,proliferation,drug resistance and apoptosis of the CD34+cells.These proteins may be the specific and or associated molecules of CD34+cells of AML-M2subtype leukemia,they may not only play important role in the leukemogenesis of AML-M2subtype leukemia,but also may be the targets for molecular diagnosis and therapy of the AML-M2subtype leukemia.