title: [Genotyping and minimal residual disease study in children with acute myeloid leukemia: preliminary results].
abstract: The aim of the paper is to present the initial results of molecular examination which was started in 2006 for children with acute myeloid leukemia. Better knowledge of biology of this disease, can result in establishing of new risk factors what allows more precise patient stratification to different therapeutic groups. Study was obtained patients until to 18 years of age treated according to AML-BFM 2004 INTERIM protocol in 14 centers of the Polish Pediatric Leukemia/Lymphoma Study Group. Mononuclear cells were collected from bone marrow on time points established according to the AML-BFM 2004 INTERIM protocol. Collected cells were isolated on Ficoll gradient, and RNA and DNA were isolated using TRIZOL reagent. To synthesize cDNA an amount of 1 mg of total RNA was used. To perform quantitative RT-PCR and RQ-PCR reactions 4 fusion gene transcripts (AML1-ETO, CBFb-MYH11, PML-RARA /subtype bcrl and bcr3/) were used according to the protocol established by Europe Against Cancer Program. An expression of WT1 gene was tested additionally. An analysis of ABL control gene was used to normalize of achieved results. Determination of duplication of FLT3 gene in DNA sample was performed with starters complementary to JM region. Genotyping was performed in 75 patients with acute myeloid leukemia so far. AML1-ETO fusion gene transcript was found in 14 patients (19%). PML-RARA (subtype bcr3) and CBFB-MYH11 gene transcripts were detected in 3 (4%) and 3 (4%) patients, respectively. Duplication of FLT3 gene was found in 4 (5.3%) cases. Between 67 tested children over expression of WT1 was present in 51 patients (76%). Analysis of MRD level in subsequent time points showed systematic decrease of number of fusion gene transcript copies and gene WT1 expression. To establish the rate of molecular marker presence in AML in children and the influence of the presence of MRD on the treatment results as well, the study has to be conducted on a larger group of patients with longer follow-up.
authors.affiliation: Klinika Onkologii i Hematologii Dzieciecej, PA Instytut Pediatrii, UJ-CM w Krakowie. balwierz@mp.pl
authors.forename: Walentyna
authors.initials: W
authors.lastname: Balwierz
authors.forename: Jacek J
authors.initials: JJ
authors.lastname: Pietrzyk
authors.forename: Gracjan
authors.initials: G
authors.lastname: Wator
authors.forename: Karol
authors.initials: K
authors.lastname: Stozek
authors.forename: Tomasz
authors.initials: T
authors.lastname: Klekawka
authors.forename: Kinga
authors.initials: K
authors.lastname: Kwiecińska
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Dłuzniewska
authors.forename: Michał
authors.initials: M
authors.lastname: Matysiak
authors.forename: Iwona
authors.initials: I
authors.lastname: Malinowska
authors.forename: Barbara
authors.initials: B
authors.lastname: Sikorska-Fic
authors.forename: Anna
authors.initials: A
authors.lastname: Balcerska
authors.forename: Lucyna
authors.initials: L
authors.lastname: Maciejka-Kapuścińska
authors.forename: Danuta
authors.initials: D
authors.lastname: Sońta-Jakimczyk
authors.forename: Renata
authors.initials: R
authors.lastname: Tomaszewska
authors.forename: Alicja
authors.initials: A
authors.lastname: Chybicka
authors.forename: Maryna
authors.initials: M
authors.lastname: Krawczuk-Rybak
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Muszyńska-Rosłan
authors.forename: Wojciech
authors.initials: W
authors.lastname: Młynarski
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Stolarska
authors.forename: Tomasz
authors.initials: T
authors.lastname: Urasiński
authors.forename: Elzbieta
authors.initials: E
authors.lastname: Kamieńska
authors.forename: Grazyna
authors.initials: G
authors.lastname: Sobol
authors.forename: Maria
authors.initials: M
authors.lastname: Wieczorek
authors.forename: Grazyna
authors.initials: G
authors.lastname: Karolczyk
authors.forename: Mariusz
authors.initials: M
authors.lastname: Wysocki
authors.forename: Sylwia
authors.initials: S
authors.lastname: Kołtan
authors.forename: Jerzy R
authors.initials: JR
authors.lastname: Kowalczyk
authors.forename: Beata
authors.initials: B
authors.lastname: Wójcik
authors.forename: Teofila
authors.initials: T
authors.lastname: Ksiazek
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Szewczyk
mesh_terms: D000293:Adolescent; D002648:Child; D002675:Child, Preschool; D005260:Female; D016162:Genes, Wilms Tumor; D005819:Genetic Markers / Q000235:genetics*; D005838:Genotype; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D018365:Neoplasm, Residual / Q000235:genetics*; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000188:drug therapy / Q000235:genetics*
chemical_list: D005819:Genetic Markers