Study Of FLT3 And DCK Gene Mutations In Acute Leukemia By Denaturing HPLC

Objective 1.Deficiency of functional deoxycytidine kinase (DCK) is a common characteristic for resistance to cytarabine (Ara-C). Mutations of DCK gene have been shown to be responsible for DCK deficiency in vitro. To investigate whether point mutatons exist in DCK gene and its significance in acute leukemia(AL), mutations in the DCK gene from 160 patients with acute leukemia were screened and the mutation site identified. 2. To detect mutation in the FLT3 (Fms-like tyrosine kinase3) gene from 116 patients with acute myelogenous leukemia (AML) and identify the mutation site. Observing the correlation between the clinical phenotype of patients with AML gene mutation.Method 1.The denaturing high-performance liquid chromatography (DHPLC) was used to detect DCK mutations of complete coding frame in 160 patients with acute leukemia. 2. Total mRNAs extracted from 116 AML patients were first analyzed for the presence of juxtamembrane length mutations by RT-PCR. Denaturing high-performance liquid chromatography (DHPLC) was introduced to screen FLT3 gene point mutation from exon 11 to exon 24. The DNA amplification fragment which DHPLC elution profiles showed different from corresponding normal elution profile were sequenced to identify the position and nature of mutation.Result1. DCK gene: Five patients were detected to bear nucleotide changes at nt523 (C/T; GenBank accession No.NM₀00788) and produced the amino acid changes...