ASXL1, DNMT3A, FLT3, KIT, NRAS, TP53 and WT1 genes mutations detection in acute myeloid leukemia with unspecified karyotype using direct sequencing technique
DOI: 10.22138/2500-0918-2016-14-4-38-51
A.V. Vinogradov, A.V. Rezaykin, D.V. Izotov, A.G. Sergeev
Sverdlovsk Regional Ministry of Health, Yekaterinburg, Russian Federation;
Ural State Medical University, Yekaterinburg, Russian Federation;
Sverdlovsk Regional Clinical Hospital N 1, Yekaterinburg, Russian Federation
Abstract.
Aim: to estimate the frequency of mutations in ASXL1, DNMT3A, FLT3, KIT, NRAS, TP53 and WT1 genes in acute myeloid leukemia (AML) patients (pts) with unspecified karyotype using direct automatic sequencing technique.Materials and methods. Bone marrow and peripheral blood samples obtained from 41 AML pts aged 21 to 82, treated in Sverdlovsk Regional Hematological Centre during the period 2008–2016. Distribution of the pts according to FAB-classification was as follows: AML M0 — 1, M1 — 3, M2 — 23, M4 — 10, M5 — 1, acute myelofibrosis — 1, AML pts with unspecified morphological variants — 2. AML pts with M3, M4eo morphologic subtype according to FAB-classification, with normal or abnormal karyotype (in G-banding), t(8;21), inv(16), t(9;22), abn11q23 (in PCR) and NPM1 gene exon 12 insertion (in sequencing) were excluded from research.
Detection of mutations in ASXL1, DNMT3A, FLT3, KIT, NRAS, TP53 and WT1 genes performed by direct sequencing. Sequencing realized using an automatic genetic analyzer ABI Prism 310. Results. The average frequency of functionally significant mutations in all investigated genes among the treated AML pts was 26.8%, and low than cumulative frequency of point mutation in AML with normal and abnormal karyotype. Average frequency of mutations in KIT gene exons 7–12 and 16–19 was 15.0%, ASXL1 gene exons 12–13 – 12.5%, DNMT3A exons 18–26 – 11.1%, WT1 gene exons 6–9 — 11.1%, FLT3 gene exons 12–15 and 19–21 —10.8%, NRAS gene exons 1–4 — 5.4%, TP53 gene exons 4–11 — 3.6%. Multiple point mutations in investigated genes detected in 9.8% AML pts with unspecified karyotype (usually KIT gene non-synonymous substitution and deletion). In 2 cases 2 different gene mutations were co-existed in samples: AML M5 with FLT3 ITD and KIT gene deletion n. Del. 1529–1774, AML M2 with WT1 gene deletion n. Del. 1289–1372 and DNMT3A gene non-synonymous substitution c. 2645G>A. In 2 another samples, both AML M2 with KIT gene non-synonymous transversion c. 1621А>С additional TP53 gene exon 6 tandem duplication and KIT gene non-synonymous transition c. 1636A>G were detected, respectively.
Keywords:
acute myeloid leukemia, unspecified karyotype, ASXL1 gene, DNMT3A gene, FLT3 gene, KIT gene, NRAS gene, TP53 gene, WT1 gene, sequencing
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Authors:
Vinogradov Alexander V.
Sverdlovsk Regional Ministry of Health, Department of Specialized and Hi-Tech Medical Care
MD, chief therapist, scientific advisor
Russian Federation, 620014, Yekaterinburg, Weiner str., 34b
a.vinogradov@egov66.ru
Rezaykin Alexey V.
Ural State Medical University
Department of Microbiology, Virology and Immunology, Laboratory of Molecular Diagnostic
MD, professor assistant
Russian Federation,620028, Yekaterinburg, Repin str., 3
alexrez@usma.ru
Izotov Denis V.
Ural State Medical University
intern
Russian Federation, 620028, Yekaterinburg, Repin str., 3
phoenus@yandex.ru
Sergeev Alexander G.
Ural State Medical University
Department of Microbiology, Virology and Immunology, Laboratory of Molecular Diagnostic
MD, professor, chief of the department, scientific chief of the laboratory
Russian Federation, 620028, Yekaterinburg, Repin str., 3
sergeev@usma.ru
Received 08.11.2016
For citation:
Vinogradov A.V., Rezaykin A.V., Izotov D.V., Sergeev A.G. ASXL1, DNMT3A, FLT3, KIT, NRAS, TP53 and WT1 genes mutations detection in acute myeloid leukemia with unspecified karyotype using direct sequencing technique [Primenenie tehnologii prjamogo avtomaticheskogo sekvenirovanija dlja detekcii mutacij genov ASXL1, DNMT3A, FLT3, KIT, NRAS, TP53 i WT1 pri ostryh mieloidnyh lejkozah c neutochnennym kariotipom] Vestn. Ural. Med. Akad. Nauki. – Journal of Ural Medical Academic Science. 2016, no. 4, pp. 38–51. DOI: 10.22138/2500-0918-2016-14-4-38-51 [In Russ.]
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