A Preliminary Investigation;Screening Of Serum Protein Markers By Protein Microarray Technology In Acute Myeloid

ObjectiveWith the identification of cell genetics, molecular biology and therapeutic target inrecent years, Stratification treatment and target treatment significantly improved the acutemyeloid leukemia (acute myeloid leukemia, AML) clinical efficacy. The detection ofcytogenetic and molecular genetic has become the main basis in clinical classification ofAML. The United States National Comprehensive Cancer Network (NCCN) and ChinaAML (not APL) treatment guidelines based on cytogenetic and molecular biologicalindicators, risk classification of AML are divided into good outcome, prognosis ofsecondary and bad outcome groups. But many patients with good prognosis are mismatchwith clinical treatment the treatment effect is poor, difficult to alleviate, after remission ofeasy recurrence. It’s related to complex of cytogenetic and molecular biology, detectionerror factors，is still a lot more complex leukemia prognostic factors did not clarify. So, weneed to explore detection method which is better comprehensive and accurate on prognosisof patients with leukemia.Protein chip is a proteomic research tools.the advantage are high throughput, highsensitivity, easy to operate and high practicality. We choose RayBiotech biotinylatedantibody chip which can detect507kinds of human protein expression levels. It includecytokines, chemokines, cytokines, growth factors, angiogenesis factor, soluble receptors,cell adhesion molecules and other proteins. The chip shows widely application value inidentification of key risk factors, screening biomarkers and target of drug. Therefore, wetest serum specimens from AML patients with different cell genetics and molecular biologyby the chip, to expect selected clinical judgment molecular prognostic value of AMLmarkers, which lays the foundation for further preparation of low density protein chip forclinical prediction prognosis of AML patients and individual chemotherapy. Method1.According to cytogenetic marker、genetic test and clinical therapeutic efficacy, totally113patients with newly diagnosed AML(non-APL) hospitalised in our department fromaugust2011to January2013were divided into good outcome、medium outcome and badoutcome group.2.The serum of11patients with newly diagnosed AML and5normal cases were testedthrough biotinylated antibody chip with detected507kinds of human protein expressionlevel.3.The serum of18AML patients (9good outcome cases and9bad outcome cases) and9normal cases were tested through ELISA.4. The result were analyzed in advance by software AAH-BLM-1and t test.Result1. According to the age, leukocyte count, cytogenetic, biological markers and clinicaltherapeutic efficacy,71cases are divided into three groups，10cases are in the goodoutcome group,27cases are in the medium outcome group，34cases In the good outcomegroup respectively.2．Compared with the5normal cases, the22signs factors are significant different in11cases of AML(P＜0.05). Growth factors contain MFG-E8, and Osteoactivin, Hepassocin,M-CSF, and M-CSFR, Insulin R. Interleukin family contain IL-20、IL-3、IL-1F8/FIL1eta、IL-2R alpha、IL-2R beta/CD122、IL-1R6/IL-1Rrp2. Apoptosis factorsis containFas/CD95, adhesionmolecular, ICAM-3/CD50, Chemokines,6Ckine/CCL21. Lipocalinprotein: Lipocalin-1, new types of transmembrane proteins, TMEFF1/Tomoregulin-1,matrix. Metalloproteinase family members: MMP-3members of the transforming growthfactor Super family, GFR alpha-4tumor necrosis factor receptor superfamily: TNF-alpha,and GITR/TNFRF18, and DR6/TNFRSF21.3. Compared with the5good outcome cases, the11signs factors are significantdifferent in6bad outcome cases (P＜0.05).Growth factors contain VEGF-D, IGFBP1.Matrix metalloproteinases inhibitors: TIMP-2. Endogenous angiogenesis inhibitors: ES.proteolytic enzymes: UPA. Chemokines: MIP-2, FGF-19. Binding proteins: LBP.Adhesionmolecules: L-selectin/CD62L. Apolipoprotein: SAA.,interleukin17receptor D.4.9good outcome cases and9bad outcome cases are tested by OPN ELISA Kit. It revealed that the serum OPN levels are significantly higher in bad outcome cases withFLT3/ITD-possitve than good outcome cases with AML1/ETO、NPM1、CEBPA-possitveand normal cases(P<0.05).The serum OPN levels are significantly higher in good outcomecases with AML1/ETO、CEBPA-possitve than normal cases(P<0.05).Conclusion1.Prognosis and risk stratification of AML are closely associated with the age of onset，first-time WBC count，cell type, cell genetics/molecular genetics, clinical treatment effects.2.The high express of22sign factor paly an important role in proliferation,differentiation and apoptosis, invasion of AML cell. It’s closely related the occurrence anddevelopment of AML.3. signs factor in11cases play an important role in extramedullary infiltration,coagulation and fibrinolysis system of regulation, hematopoietic reconstruction afterhematopoietic stem cell transplantation, which is high expression in bad outcome group ofAML. Available as the basis to build new AML prognosis index system，it provides a newmethod in prognosis and the individualized treatment in AML.4. The OPN expression of bad outcome group in AML with of FLT3/ITD-positive issignificantly higher than good outcome group. It confirmed that OPN is unfavorableprognostic factors in AML. It also proved that the stability of serum specimens in AML andthe reliability of the measured protein chip.