Research Of CEBPA And FLT3-ITD Gene Mutations In Patients With Acute Myeloid Leukemia

Objectives To detect the mutations of CCAAT/enhancer binding protein-alpha (CEBPA) and Internal tandem duplication of the FLT3 (FLT3-ITD)gene in acute myeloid leukemia (AML), observe the distribution of CEBPA and FLT3-ITD mutations in different subsets of AML, explore the relation of CEBPA and FLT3-ITD mutations with regard to clinical feature of AML, so that to provide theory evidence for clinic diagnosis, prognosis evaluation and targeted therapy.Methods1.CEBPA mutations were detected using ?uorescence-based Multiplex-PCR coupled with capillary electrophoresis and ?uorescence detection in 85 patient samples.2.All patient samples with differences in amplicon length were subjected to nucleotide sequencing.3. FLT3-ITD mutations were determined by agarose electrophoresis of genomic DNA-PCR products.Resultes1. CEBPA mutations were detected in 7 0f 85 (8.2%) AML cases. In acute myeloid leukemia with normal karyotype (AML-nk), CEBPA mutations were detected in 4 0f 46 (8.7%) AML cases.2. 10 CEBPA mutations were detected by sequencing in 7 patient samples with differences in amplicon length,and no mutation detected in 10 without differences in amplicon length. 3 patients had mutations occurred in both N-terminal part and basic-leucine zipper (bZIP) domain, 1 had an N-terminal frameshift mutation and the remaining 3 had an inframe insertion or deletion in the bZIP domain. 5 mutations were not reported before : nt1503₁505delGCA, nt1643₁648delTCAAG,nt792₇93insCTAC, nt1535₁536insCAC and nt760 subG>T。3. 17 of 85(20.0%) AML cases were detected with FLT3-ITD mutations. 19.5% of AML-nk were FLT3 ITD-AML. Accoding to the electrophoresis results, the length of ITD was variety.4. FLT3-ITD was detected in 2 of 7(28.6 %) patients with AML who had CEBPA mutations.5. Aggregate analysis CEBPA and FLT3-ITD gene mutations in AML cases, In the CEBPA-mutated/FLT3 ITD- mutated group, 2 patients were conducted as induction therapy, patients reached complete remission(CR), the CR rate was 100.0%(2/2). Conclusions1. CEBPA mutations had been found in AML with an incidence of 8.2% in adults, while FLT3-ITD mutations had been found of 20.0% in patients with acute myeloid leukemia. FLT3-ITD was detected in 28.6 % patients with AML who had CEBPA mutations. 10 different variant mutations have been identified in this research, 5 had been reported, 5 were newly found to date.2.The method of ?uorescence-based Multiplex-PCR to detected CEBPA was very easy, fast and cheap,and it may be used as a routine molecule detection technique in clinic.3. To detect CEBPA and FLT3-ITD mutations could provide theory evidence for risk-adapted treatment protocols to improve treatment outcome and earlier period direct interference or target therapy with AML.