Molecular genetic alterations in acute myeloid leukemia.

Cytogenetic and molecular genetic analyses are necessary for precise assessment of diagnosis, prognosis and treatment of patients with AML. The karyotypic analysis allows the distribution of patients into the basic risk groups, while the methods of molecular biology offer further possibilities to stratify patients within particular risk subgroups. Moreover, using quantitative PCR, they enable to follow the course of minimal residual disease (MRD) and foresee the eventual relapse of the disease. The aim of this thesis was to analyse the prognostic impact of new molecular markers in patients with AML, particularly in those with favourable (acute promyelocytic leukemia (APL), CBF-AML) and intermediate (influence of FLT3 mutations and others) cytogenetic profiles. The presence of fusion genes PML/RARα, AML1/ETO and CBFβ/MYH11 was tested by qualitative PCR. Patients harbouring fusion genes AML1/ETO or CBFβ/MYH11 (CBF-AML) were further analysed using either sequencing or restriction digest analysis, for the presence of C-KIT, K-RAS, N-RAS and FLT3 mutations. Patients with intermediate cytogenetic risk were tested for presence of internal tandem duplications of FLT3 (FLT3/ITD), mutations in tyrosine kinase domain of FLT3 (FLT3/TKD), DNMT3A and ASXL1 mutations. Cases with a complex karyotype were screened..