| Objective:To screen the FMS-like tyrosine kinase 3 internal tandem duplication (FLT3-ITD) mutation in patients with acute myeloid leukemia (AML), and to study the relationship between their clinical characteristics and the FLT3-ITD mutation, as well as the impact of FLT3-ITD mutation on prognosis.Methods:284 newly diagnosed AML patients were enrolled. Mononuclear cells were isolated from their bone marrow aspirates, and then genomic DNA was extracted. The region spanning exonl4 and 15 of FLT3-ITD gene was amplified using polymerase chain reaction method; electrophoresis of the PCR products was performed and then followed by gel imaging. The morphology, cytogenetic characters and other clinical characteristics of these AML patients were compared between the FLT3-ITD mutated patients and wild type patients. The prognostic impact of FLT3-ITD mutation was also evaluated.Results:FLT3-ITD mutation was detected in 58 (20.4%) of the 284 AML patients. The morphology distribution was 13 cases in M1,10 cases in M2,11 cases in M 3, and 24 cases in M5. FLT3-ITD mutation was detected in 24 (24.5%) of the 98 AML patients with normal karyotype, the detection rate was significantly higher than in other AML karyotypes (p<0.05). Compared with FLT3-ITD wild type patients, the FLT3-ITD mutated patients had higher white blood cell count at diagnosis (p<0.05), but there was no difference in other clinical characteristics, including age, gender, hemoglobin level, platelet count and progenitor cell percentage in bone marrow. The CR rate of FLT3-ITD mutated patients was 61%, which was lower than that of the FLT3-ITD wild type patients(p<0.05); the median survival of FLT3-ITD mutated patients was 7 months, the overall survival was shorter than the FLT3-ITD wild type patients(p<0.05).Conclusion:FLT3-ITD was a common genetic mutation in AML patients, especially in patients with normal katyotype. FLT3-ITD mutated patients had significant inferior survival, indicating FLT3-ITD mutation had negative impact on survival. Objective:To screen the Nucleoplasmin (NPM1) mutation in patients with acute myeloid leukemia (AML), and to study the relationship between their clinical characteristics and the NPM1 mutation, as well as the impact ofNPMl mutation on prognosis.Methods:284 newly diagnosed AML patients were enrolled. Mononuclear cells were isolated from their bone marrow aspirates, and then genomic DNA was extracted. The region spanning exon12 of NPM1 gene was amplified using polymerase chain reaction method and then subjected to sequencing. The PCR product of mutated patient was used to construct NPM1 plasmid standard, the plasmid standard of internal reference gene ALB was also constructed. Real-time quantitative PCR (Q-PCR) was employed to detect the fluorescence of the PCR products from the serial diluted plasmid standards. The standard curve was then illustrated for NPM1 and ALB gene. The same Q-PCR system was used to screen the NPM1 mutation in the patients' DNA samples. The morphology, cytogenetic characters and other clinical characteristics of these AML patients were compared between the NPM1 mutated patients and wild type patients. The prognostic impact of NPM1 mutation was also evaluated.Results:NPM1 mutation was detected in 51 (18.0%) of the 284 AML patients.46 of them were mutation type A,3 were mutation type B, and 2 were mutation type D. The morphology distribution was 13 cases in M1,6 cases in M2,3 cases in M 3,1 case inM4, and 28 cases in M5. NPM1 mutation was detected in 29 (29.6%) of the 98 AML patients with normal karyotype, the detection rate was significantly higher than in other AML karyotypes (p<0.001). Compared with NPM1 wild type patients, the NPM1 mutated patients had higher white blood cell count at diagnosis, older in age, lower CD34 expression and higher progenitor cell percentage in bone marrow (all p<0.05), but there was no difference in other clinical characteristics, including gender, hemoglobin level, and platelet count. The CR rate of NPM1 mutated patients was 88.2%, the median survival of NPM1 mutated patients was 13 months, there was no dfference compared with the NPM1 wild type patients (p>0.05). However in patients with normal karyotype, when we define the patients into the NPM1+/FLT3-ITD-,NPM1+/FLT3-ITD+,NPM1-/FLT3-ITD-å’ŒNPM1/FLT3-ITD+ four groups, the difference of CR rate and survival was significant (p<0.05).Conclusion:NPM1 was a common genetic mutation in AML patients, especially in patients with normal katyotype. NPM1 mutated patients had significant superior survival, indicating NPM1 mutation had favorable impact on survival. Objective:To screen the CCAAT/enhancer binding protein a(CEBPa) mutation in patients with acute myeloid leukemia (AML), and to study the relationship between their clinical characteristics and the CEBPa mutation, as well as the impact of CEBPa mutation on prognosis.Methods:284 newly diagnosed AML patients were enrolled. Mononuclear cells were isolated from their bone marrow aspirates, and then genomic DNA was extracted. The CEBPa gene was divided into 2 parts for polymerase chain reaction amplification. The PCR product was then subjected to sequencing. For the samples whose sequencing results were hard to interpret, PCR was performed again and the PCR products were constructed into plasmid. The plasmid was transfected into competent cells, and the positive clones were picked out for sequencing. The morphology, cytogenetic characters and other clinical characteristics of these AML patients were compared between the CEBPa mutated patients and wild type patients. The prognostic impact of CEBPa mutation was also evaluated.Results:CEBPa mutation was detected in 40 (14.4%) of the 284 AML patients, including 19 cases of double mutation and 21 cases of single mutation. The morphology distribution was 1 cases in M1,22 cases in M2,1 cases in M 3,1 case in M 4, and 9 cases in M5. CEBPa mutation was detected in 30 (30.6%) of the 98 AML patients with normal karyotype, the detection rate was significantly higher than in other AML karyotypes (p<0.001). Compared with CEBPa wild type patients, the CEBPa mutated patients had lower white blood cell count at diagnosis (p<0.05), but there was no difference in other clinical characteristics, including age, gender, hemoglobin level, platelet count and progenitor cell percentage in bone marrow. The CR rate of patients with CEBPa double mutation was 91.7%, The CR rate of patients with CEBPa single mutation was 82.7%, there were no differences between CEBPa mutated and wild type patients(p<0.05). The median survival of normal karyotype patients with CEBPa double mutation was 19 months, the median survival of patients with CEBPa single mutation was 15 months. The overall survival was longer than the CEBPa wild type patients(p<0.05).Conclusion:CEBPa gene mutation was mainly detected in AML patients with M2 subtype. Only those with CEBPa double mutation had favorable prognosis. Objective:To screen the c-kit mutation in patients with core binding factor acute myeloid leukemia (CBF AML), and to study the relationship between their clinical characteristics and the c-kit mutation, as well as the impact of c-kit mutation on prognosis.Methods:284 newly diagnosed AML patients were enrolled. CBF AML patients were screened out by cytogenetic and fusion gene studies. Mononuclear cells of the patient samples were isolated from their bone marrow aspirates, and then genomic DNA was extracted. Polymerase chain reaction was used to amplify the exon 8 and exon 17 of the c-kit gene. The PCR product was then subjected to sequencing. The morphology, cytogenetic characters and other clinical characteristics of these AML patients were compared between the c-kit mutated patients and wild type patients. The prognostic impact of c-kit mutation on CBF AML was also evaluated.Results:45 of the 284 AML patients were CBF AML.c-kit mutation was detected in 17 (37.7%) of them There was significant relationship between c-kit gene mutation and CBF AML. The mutation type included:6 cases of exon 8 mutation,4 cases of D816V mutation, 2 cases of D816Y mutation,2 cases of D816H mutation and 3 cases of N822K mutation. The morphology distribution was 14 cases in M2,1 case in M 4, and 2 cases in M5. Compared with c-kit wild type patients, the c-kit mutated patients were younger in age(p<0.05), but there was no difference in other clinical characteristics, including gender, white blood cell count, hemoglobin level, platelet count and progenitor cell percentage in bone marrow. The CR rate ofpatients with c-kit mutation was%, which was similar with that of the c-kit wild type patients (p>0.05); The median survival of patients with c-kit double mutation was months, the overall survival was shorter than the c-kit wild type CBF AML patients (p<0.05).Conclusion:c-kit gene mutation is a characteristic mutation of CBF AML patients. c-kit mutated patients had significant inferior survival, indicating c-kit mutation had negativ impact on survival of CBF AMLpatients. |
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