The Differential Proteomics Analysis Of Cells From Children With Acute Lympho Blastic Leukemia

Objective:To acquire the different proteins expressed between the cells of the initial diagnosis acute lymphocytic leukemia(ALL) childrens and the healthy childrens by differencial proteomics, so as to provide basis for reaserch the developmental mechanism of ALL deeply and explore new diagnosis and cure pathway.Object and methods:1. Collect bone marrow cells of ALL childrens for 15 cases classified as experimental group and peripheral blood white cells of healthy children for 15 cases classified as normal controls.All of these ALL cases are made a definite diagnosis by bone marrow cytology review and immunologicaltype,without no more department hyperplasia.2. Obtain and splitting individual nucleus cells by the lymphocyte separation fluid and cell cracking liquid and acquire all of the protein in cells. Separate these proteins by the method of 2-DE. Guarantee 3 independent repeated experiments for every case.3. Scan gel image by transmission mode. Analyse the difference protein points by ImageMaster 2D Platinum 7.0 Training and set software testing and evaluation standards as Volume ratio>2 and anova P<0.05, acquire the different proteins expressed between two groups.4. Cut the protein spots by hand and obtain protein peptides by enzymolysis, analyse and identify the peptides by mass spectrometry with MALDI-TOF-MS and the database of IPI human human v3.07.Rrsults:1. The number of protein spots detected in experimental group and normal controls are (889±52) and (834±41) respectively,acquire 4 higher expression proteins and 11 lower expression proteins and most of their isoelectric points distribute among pH4.5--7,and molecular masses distribute among 18.4～45KD.2. Obtaine 2 proteins higher expression(Glutathione S-transferase P,Prohibitin) and 6 proteins lower expression (60S acidic ribosomal protein P0,Pyridoxine-5'-phosphate oxidase,Triosephosphate isomerase 1 variant,Peroxiredoxin 4,Actin cytoplasmic,Hypothetical protein FLJ26567) in childhood ALL cells by mass spectrometry and database.Conclusions:1. There are difference of proteins expression between ALL children and healthy children. We obtain 8 kinds of proteins and all of them are linked to the happening and development mechanism of child ALL probably.Among of them, the relationship of Glutathione S-transferase P, Actin cytoplasmic and children ALL have the similar results as previouse not proteomics researchs,and the relationship of Prohibitin,60S acidic ribosomal protein P0,Pyridoxine-5'-phosphate oxidase,Triosephosphate isomerase 1 variant,Peroxiredoxin 4,Hypothetical protein FLJ26567 and children ALL are first discovered at home and abroad.2. All of the 8 protains have the possibility to be the diagnosis markers of children ALL.When the level of Glutathione S-transferase P and Actin cytoplasmic rise and the level of 60S acidic ribosomal protein P0,Pyridoxine-5'-phosphate oxidase,Triosephosphate isomerase 1 variant,Peroxiredoxin 4,Actin cytoplasmic and Hypothetical protein FLJ26567 reduce in childen cells,it tells us to watch out for the children acute leukemia.