The Clinical Significance Of The Expression Of C-fes Gene In Leukemia Patients

Objective:The acute leukemia is one of the most common hematopoietic malignances,but the pathogenetic has not yet clear-out.Now cell differentiation can be seen as a much more predominant theory. It presumed that genetic mutation could prevent terminal differentiation of cells and cause cumulate of the immature cells to induce canceration.C-fes belongs to src family,it was dicoveried from feline sarcoma virus by Snyder and Theilen in 1969. The cellular fes gene localization is 15q25-26,encodes a 93-kilodalton protein-tyrosine kinase (Fes), c-fes mRNA is expressed in primary acute myelogenous leukemia (AML),CML,certain lymphoid malignancies,hematopoietic stem cell,CD34+ progenitor cell,vascular endothelial cell,central nervous system and leukemic cell lines such as HL-60,KG-1, TF-1, THP-1 and U937.Hematopoietic stem/progenitor cells express low levels of c-fes mRNA,but among differentiated cells, its expression is restricted to the myeloid lineages,we could not detect its expression in mature lymphocyte.Moreover, c-fes mRNA protein levels and kinase activity increase upon induction of granulocyte cell differentiation. In addition,Fes has been implicated in signal transduction pathways for several hematopoietic cytokines. This experiment intends to research the expression of c-fes mRNA in different types of leukemia and the clinical prognosis,so as to provide a new method for leukemia therapy.Methods: The c-fes mRNA was measured in 121 cases with acute and chronic leukemia ,among total there were 66 cases with AML(M1 4 cases,M2 26 cases(AML1/ETO positive 8 cases),M3 14 cases(PML/RARαpositive 9 cases),M 4 12 cases,M5 8 cases,M6 2 cases),28 cases with ALL(L1 6 cases,L2 20 cases,L3 2 cases),23 cases with CML,include 10 cases of CML-CP,6 cases of CML-AP,7 cases of CML-BP,4 cases of CLL.There were 69 cases were male and 52 cases were female. Take 20 normal cases as controls. The expression levels of c-fes mRNA were measured by real time-quantify reverse transcription polymerase chain reaction(RQ-PCR). Then approach the expression of c-fes mRNA among leukemia types and cell differentiation,so as to provide assistance in clinical therapy. HL-60 cells were cultured in RPMI 1640 and treated with different concentrations of As2O3.The status of cells proliferate were observed under microscops,the cells viability were oberved by MTT test,the differentiation of HL-60 cells were examined by NBT reduction reaction,cells were stained with Wright-Giemsa for general morphology. Flow cytometry (FCM) was used to detect the expresstion of CD11b. The expression levels of c-fes mRNA in trial groups and control group were measured by RQ-PCR.Results: 1.The expression level of c-fes mRNA in AML patients was significant higher than that of normal controls (P<0.001),There were no significant difference about expression level of c-fes mRNA between samples of ALL and normal controls (P>0.05). The expression level of c-fes mRNA in AML patients was much more higher than that of ALL patients (P<0.001).2.The expression level of c-fes mRNA in CML-CP patients was significant higher than that of normal controls (P<0.01), The expression level of c-fes mRNA in CML-AP patients was significant higher than that of control group (P<0.05),There was no significant difference of c-fes mRNA between CML-BP patients and normal controls (P>0.05). The expression level of c-fes mRNA in CML-CP patients was much more higher than that of CML-AP patients(P<0.05). The expression level of c-fes mRNA in CML-CP patients was much more higher than that of CML-BP patients(P<0.01). The expression level of c-fes mRNA in CML-AP patients was higher than that of CML-BP patients(P<0.05).The expression level of c-fes mRNA decreased or disapeared with degression of granulocyte cell differentiation.3.The patients who had higher expression level of c-fes mRNA had good prognosis,the difference had statistical significance (P<0.01),so the expression level of c-fes mRNA could help evaluating the clinical prognosis. 4.Treated with low concentrations of As2O3(0.1,0.5,1.0,2.5μmol/L), HL-60 cells were significantly differentiated and in dose-dependent and time-dependent manner.5.The cell morphology appeared to maturated after treated with low concentrations of As2O3 ( 0.1,0.5,1.0,2.5μmol/L),showing the volume grow downwards, nucleolus almost disappearance,karyoplasmic ratio diminished, karyomorphism irregular; more vacuole in intracytoplasm.6.As the RQ-PCR showed that the expression level of c-fes mRNA was up-regulated by low concentrations of As2O3(0.1,0.5,1.0,2.5μmol/L).Conclusions:1.The expression level of c-fes mRNA in AML patients and some CML patients is much higher than that of ALL patients,so the c-fes gene could be used to clarify types of leukemia.2.The patients who have high expression level of c-fes mRNA are much more sensitive to chemical therapy and have good prognosis.So the expression level of c-fes mRNA could help evaluating the clinical prognosis.3.The expression level of c-fes mRNA in CML-CP patients is higher than that of CML-AP and CML-BP patients,it means c-fes gene plays a role in cell differentiation.4.The HL-60 cells are differentiated after treated with As2O3, and it is in dose-dependent and time-dependent manner.5.The expression level of c-fes mRNA is up-regulated by low concentrations of As2O3.We could raise the expression level of c-fes mRNA so as to make the cell differented,it could be a target of therapy about leukemia in the future.