Identification of FLT3 and NPM1 Mutations in Patients with Acute Myeloid Leukaemia

Yusoff, Yuslina Mat; Seman, Zahidah Abu; Othman, Norodiyah; Kamaluddin, Nor Rizan; Esa, Ezalia; Zulkiply, Nor Amalina; Abdullah, Julia; Zakaria, Zubaidah

doi:10.31557/apjcp.2019.20.6.1749

Cited by 10 publications

(12 citation statements)

References 38 publications

(25 reference statements)

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“…Yusoff et al said that the most common genetic abnormality in AML were NPM1 mutations at exon 12 and he found it in approximately 24 to 45% of all AML cases. 11 Gallagher in his study found that more than 50% of normal karyotype-AML cases harbor mutations in the NPM1 gene, and he concluded that it the highest incidence of any mutation in AML. 12 Mencia-Trinchant, et al found that half of his patients with AML have normal cytogenetics whereas 40% to 50% of these patients have NPM1 mutations.…”

Section: Discussionmentioning

confidence: 99%

Molecular study of Nucleophosmin 1(NPM1) gene in acute myeloid leukemia in Kurdish population

Othman¹,

Mohammad²,

Saeed³

2021

Afr H. Sci.

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Background: In patients with Acute Myeloid Leukemia (AML) the most frequent acquired molecular abnormalities and important prognostic indicators is nucleophosmin-1 (NPM1) mutations. Our study aims was molecular study of Nucleop- hosmin -1 gene in Acute Myeloid Leukemia in Kurdish population. Patients &Methods: A total of 50 patients with AML, (36) of them attended Nanakaly Hospital and (14) attended Hiwa Hospital and 30 healthy subjects as control were selected randomly, all were matched of age and gender. Polymerase chain reaction (PCR) was used for detection of NPM1 gene mutation. Three samples of PCR product for NPM1 gene mutations were sequenced, and mutations were determined by comparison with the normal NPM1 sequence NCBI (GenBank acces- sion number NM_002520). Results: Out of 50 patients with AML, 5 (10%) of them were NPM1 gene mutation positive, and 45 (90%) were negative. The mutation were a base substitution (C to A), (G to C), (G to T), transversion mutation in addition of frame shift mutation and all mutated cases were heterozygous and retained a wild type allele. Conclusion: Identification of NPM1 mutations in AML are important for prognostication, treatment decision and optimi- zation of patient care. Keywords: Acute myeloid leukemia; Nucleophosmin-1 (NPM-1) gene mutation; PCR.

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Section: Discussionmentioning

confidence: 99%

Molecular study of Nucleophosmin 1(NPM1) gene in acute myeloid leukemia in Kurdish population

Othman¹,

Mohammad²,

Saeed³

2021

Afr H. Sci.

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“…For FLT3-ITD mutation detection, the primer set used included forward primer 5'-GCA ATT TAG GTA TGA AAG CCA GC -3' (ITD_14F) and reverse primer 5'-CTT TCA GCA TTT TGA CGG CAA CC -3' (ITD_15R) (Integrated DNA Technologies, USA). 14 Polymerase chain reaction (PCR) assay was performed in a total volume of 25 µL of 12.5µL HotStarTaq master mix (Qiagen, Germany), 1 µL (10 µM) of forward and reverse primers respectively, 8.5 µL nuclease-free water, and 2 µL (10 ng/µL) genomic DNA. The PCR conditions included initial denaturation at 95°C for 7 minutes and followed by 35 cycles of denaturation (at 94°C for 1 minute), annealing (at 58°C for 45 seconds), and extension (at 72°C for 1 minute).…”

Section: Flt3-itd Mutation Detectionmentioning

confidence: 99%

“…The primer set utilised for FLT3-D835 mutational analysis was 5'-CCG CCA GGA ACG TGC TTG-3' (D835_F) as the forward primer and 5'-GCA GCC TCA CAT TGC CCC-3' (D835_R) as the reverse primer (Integrated DNA Technologies, USA). 14 The total volume used for the PCR assay was 25 µL, including 12.5 µL Hot Star Taq master mix (Qiagen, Germany), 1 µL (10 mM) of forward and reverse primers respectively, 2.5 µL Q-Solution (Qiagen, Germany), 6 µL nuclease-free water and 2 µL DNA template (10 ng/ml). The PCR conditions were initial denaturation at 95°C for 9 minutes followed by 35 cycles of denaturation (at 94°C for 18 seconds), annealing (at 59°C for 1 minute), and extension (at 72°C for 1 minute).…”

Section: Flt3-d835 Mutation Detectionmentioning

confidence: 99%

“…The primer set used included 5'-TTA ACT CTC TGG TGG TAG AAT GAA-3' (NPM1_F) and 5'-TGT TAC AGA AAT GAA ATA AGA CGG-3' (NPM1_R) (Integrated DNA Technologies, USA). 14 The PCR cycle protocol included initial denaturation at 94°C for 3 minutes followed by 35 cycles of denaturation (at 95°C for 1 minute), annealing (at 58°C for 27 seconds), and extension (at 72°C for 2 minutes). The final extension was also carried out at 72°C, for 7 minutes (C1000 Touch Thermal Cycler, BIO-RAD, Singapore).…”

Section: Npm 1 Mutation Detectionmentioning

confidence: 99%

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Detection of FMS-Like Tyrosine Kinase 3 (FLT3) and Nucleophosmin 1 (NPM1) Mutations from Marrow Tissues in Patients with Acute Myeloid Leukaemia

Hamdan

Mansoor

Muhammad

et al. 2022

imjm

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INTRODUCTION: Acute myeloid leukaemia is a haematological malignancy with diverse cytogenetic abnormalities and molecular mutations. Amongst the important mutations are FMS-related tyrosine kinase 3 (FLT3) and nucleophosmin 1 (NPM1) gene mutations. These mutations have been shown to be of prognostic significance. A cross-sectional study to examine the frequency of these mutations and their association with the haematological and cytogenetic characteristics of the cases was carried out in Kuantan, Pahang, Malaysia. MATERIALS AND METHODS: A total of 43 cases were included in the study. Polymerase chain reaction-based assays were employed for mutation detection from the retrieved trephine biopsy tissue blocks. Mutation positivity was subsequently validated by Sanger DNA sequencing. RESULTS: Six of the 43 cases (14.0%) of the acute myeloid leukaemia were positive for FLT3-type internal tandem duplications (FLT3-ITD) and a similar proportion (6/43, 14.0%) were positive for NPM1 mutations. FLT3 mutations at codon D835 (FLT3-D835) mutation was identified in three of the cases (7.0%) while concurrent mutations of NPM1 and FLT3-ITD were seen in two of the mutation-positive cases (4.7%). The total white cell count was found to be significantly higher in patients with FLT3 mutations (p=0.001). Other haematological parameters and the cytogenetic results did not reveal any significant association with the mutational status. CONCLUSION: The frequency of FLT3-ITD, FLT3-D835, and NPM1 mutations among AML patients were 14%, 7%, and 14% respectively. Follow-up studies to include the clinical parameters and the treatment outcomes are advocated.

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“…Previous studies have revealed acute myeloid leukemia (AML) as a clinical and biological highly heterogeneous clonal disease that considered originating from hematopoietic stem cells, which characterized by impaired myelopoiesis and blast proliferation (Estey and Döhner, 2006;Saultz and Garzon, 2016). Genome-wide studies using next generation sequencing (NGS), direct sequencing and high-resolution single nucleotide polymorphism (SNP) array have been provided major insight into the panel of the prognostic markers of AML (Liersch et al, 2014;Mat Yusoff et al, 2019). These markers, which encompass genetic mutations, constitute the adoption of risk stratification and therapeutic decision-making in the overall survival of AML patients (De Kouchkovsky and Abdul-Hay, 2016;Kansal, 2016).…”

Section: Introductionmentioning

confidence: 99%

Detection of TET2 Mutation in Patients with De Novo Acute Myeloid Leukemia: A Mutation Analysis of 51 Iranian Patients

Chehreghani

Sadeghian

Ayatollahi

et al. 2022

Asian Pac J Cancer Prev

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Acute myeloid leukemia (AML) is a heterogeneous clonal disease that is considered to originate from hematopoietic stem cells, which are characterized by impaired myelopoiesis and blast proliferation. TET oncogene family member 2 (TET2) mutations are frequent in myeloid malignancies and several studies have assessed the clinical importance of TET2 mutations. However, its frequency ratio has not yet been fully clarified. Method: Hence, our study was aimed to analyze TET2mut in patients with de-novo AML and their association with clinical, molecular characteristics and Nucleophosmin 1 (NPM1), Fms-like tyrosine kinase 3 (FLT3), CCAAT Enhancer Binding Protein Alpha (CEBPA) and Wilms’ tumor protein ( WT1 ) gene expression. Fifty-one Iranian patients were screened by polymerase chain reaction (PCR) and direct sequencing to evaluate TET2 mutations frequency. Results: Out of all patients, 10 mutations in 8 patients (15.6%) were detected and closely associated with higher age and higher hemoglobin levels (p-value <0.05). Although FLT3, NPM1 and CEBPA gene expression did not show any significant correlation with TET2mut, cytogenetically normal acute myeloid leukemia (CN-AML) patients appear to bear TET2mut more frequently with lower platelet counts. Monocyte-lineages leukemia has seemed to be more linked with TET2mut in these patients. Conclusion: Our study suggests the frequency of TET2mut in our study (15.6%) is in line with previous studies and reveals the critical role of TET2 in myeloid transformation, especially in leukemia with monocytic subtypes.

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Identification of FLT3 and NPM1 Mutations in Patients with Acute Myeloid Leukaemia

Cited by 10 publications

References 38 publications

Molecular study of Nucleophosmin 1(NPM1) gene in acute myeloid leukemia in Kurdish population

Molecular study of Nucleophosmin 1(NPM1) gene in acute myeloid leukemia in Kurdish population

Detection of FMS-Like Tyrosine Kinase 3 (FLT3) and Nucleophosmin 1 (NPM1) Mutations from Marrow Tissues in Patients with Acute Myeloid Leukaemia

Detection of TET2 Mutation in Patients with De Novo Acute Myeloid Leukemia: A Mutation Analysis of 51 Iranian Patients

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