The Expression Of Serum UPA Is Related To The Prognosis Of Acute Myeloid Leukemia And A Preliminary Investigation On The Related Signaling Pathway

Background and objectiveRefractory acute myeloid leukemia(AML)is still a difficult problem in the clinical treatment.During the course of AML treatment,a large number of normal blood cells would be inhibited when all kinds of chemotherapy drugs with definite curative effect kill the leukemia cells.And the risk of bleeding and infection are increased,which even lead to death.Moreover,in order to evaluate the response of patients to treatment and the effect of chemotherapy,repeated bone marrow aspiration should be performed in AML patients.Bone marrow puncture is an invasive operation which undoubtedly increase the pain of patients.Therefore,to bring maximum benefits to AML patients,constantly exploring new targeted therapy and serum prognostic indicators has become an urgent problem to be solved in the following research work.The urokinase plasminogen activator(u PA)is a serine protease secreted as zymogen(pro-urokinase).It is activated after binding to its receptor,u PAR(also called CD87).u PA is involved in the occurrence of extramedullary infiltrations and hemorrhage complications.But it is still not clear whether the u PA protein is associated with the prognosis of AML patients.Ostcopontin(OPN)is a chemokine rich in sialic acid.It has been linked to the occurrence and development of tumor by binding to CD44,alpha V beta 3(?V?3)and other cell surface receptors.In solid tumors,OPN increase the expression of u PA protein by activating the?v?3 /PI3K/NF-kappa B(NF-?B)pathway,which can further promote the invasion and metastasis of tumor cells,tumor angiogenesis,chemotherapy resistance and so on.In AML,whether OPN also perform its role in promoting the expression of u PA protein? And how dose this effect realize?In the following study,our research group will determine the level of u PA protein in the serum of AML patients with different prognostic stratification before treatment in the hope of bringing a new serological indicator for AML patients.To study the relationship between OPN and u PA protein in AML cells and the related signaling pathway will contribute to bring new ideas and important theoretical basis for the targeted therapy of AML patients.Methods1.To investigate the relationship between the expression of serum u PA protein and the prognosis of AML1.1 Collect the serum from newly diagnosed AML(non APL)and the healthy control group,then stratify the AML patients into different groups according to cell genetics,chromosome karyotype and other related prognostic indicators;1.2 Serum u PA protein level of some untreated AML patients with different prognosis were determined by enzyme linked immunosorbent assay(ELISA);1.3 Analyze and compare the data obtained.2.To study the relationship between OPN and u PA protein in refractory AML cells and to preliminarily study the related signaling pathway2.1 Culture the mv4-11 leukemia cell lines characteristic of the mutation of FLT3-ITD positive.Dispose these cells with different concentrations of recombinant human osteopontin(rh OPN).Western blot was applied to detect the expression of u PA protein2.2 Pretreat the cells with PI3 K inhibitor or NF-?B inhibitor and then process with rh OPN.Western blot was applied to detect the expression of u PA protein in each group.2.3 Analyze the gray value of the strip by using J Image software.The data were analyzed by SPSS 18 softwareResults1.The expression of serum u PA protein was significantly increased in AML patients than the healthy subjects(P < 0.05).A significant increase of serum u PA levels was found in poor prognosis AML patients compared to other control groups(all P values< 0.05).2.With increasing the concentrations of recombinant human osteopontin(rh OPN),the expression of u PA protein was increased.In the inhibitor groups,the expression of u PA protein was reduced with the increase dose of PI3 K inhibitor or NF-?B inhibitor.Conclusion1.The expression of u PA has a close relationship with the risk stratification of AML.Monitoring the serum u PA protein level contributes to a better judgment of the prognosis for newly diagnosed patients and hence guiding individual treatment.2.OPN can contribute to the expression of u PA.The inhibitor of PI3 K and NF-?B could reduce the expression of u PA protein induced by OPN.PI3 K and NF-?B are the key molecules in this signaling pathway.