Adolescent ; Adult ; Female ; Fusion Proteins, bcr-abl ; genetics ; Gene Expression ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Leukemia, Myeloid ; genetics ; Male ; Oncogene Proteins, Fusion ; genetics ; Retrospective Studies

OBJECTIVETo explore the clinical and laboratory characteristics in favor of the diagnosis of Ph/BCR-ABL positive acute myeloid leukemia (Ph/BCR-ABL⁺ AML).

METHODSRetrospectively analyzed the clinical and laboratory characteristics of 12 Ph/BCR-ABL⁺ AML cases from Feb, 2006 to Dec, 2013, with classic myeloid blast crisis of chronic myeloid leukemia (CML-MBC) as control, and followed-up the survival in these two cohorts of patients.

RESULTSThe median age of 12 Ph/BCR-ABL⁺ AML was 27.5 years, 10 cases (83.3%) showed non/mild splenomegaly, and mainly comprised of M₂ and M₄ subtypes according to FAB classification. The median number of basophils and megakaryocytes in peripheral blood and bone marrow was lower than that of CML-CBC patients. All the cases expressed myeloid antigens, 8 cases (66.7%) expressed CD34, 11 cases were detected with t(9;22), 5 cases (45.5%) with additional chromosomal abnormalities, including 1 case of inv(16). All the cases had BCR-ABL transcripts at diagnosis:3(25.0%) cases were e1a2 type and the remaining was b2a2/b3a2type, among which 1 case coexpressed CBFβ-MYH11. Two out of 6 cases existed AML-like mutations:1 case of CEBPA and the other of FLT3-TKD. For all the patients, 7 cases achieved complete remission (CR), including 6 out of 7 cases receiving induction chemotherapy combined with tyrosine kinase inhibitor (TKI) achieved CR, and 1 out of 3 cases receiving chemotherapy alone achieved CR. The median overall survival was 16.5 months, that of allo-HSCT group was 33.5 months, which was higher than that of non-HSCT group (5.5 months).

CONCLUSIONThe expression of e1a2 type BCR-ABL, the coexpression of fusion genes which were more common in AML, the existence of AML-like mutations were all indications of a de novo Ph/BCR-ABL⁺ AML. Low induction CR rate and short survival of Ph/BCR-ABL⁺ AML implied that chemotherapy combined with TKI and followed by allo-HSCT in CR was the only effective way to improve their survival.

Adult ; Blast Crisis ; Chromosome Aberrations ; Fusion Proteins, bcr-abl ; Humans ; Leukemia, Myeloid, Acute ; Oncogene Proteins, Fusion ; Protein Kinase Inhibitors ; Retrospective Studies

Objective: To investigate whether fusion protein SD-HA could regulate its downstream signaling molecule activity by competing with the phospho-BCR-ABL Y177 site, and its mechanisms to inhibit proliferation and induce apoptosis of K562 cells. Methods: Co-immunoprecipitation interaction technology analysis of fusion protein SD-HA functioned by potently binding to the phospho-BCR-ABL Y177 site, Ras, MAPK and Akt activities were observed in the Ad5F35-SD-HA-treated cells. Western blot analyses of SD-HA fusion protein on cell membrane receptor pathway to death cascade caspase-8, caspase-3 and PRAP were performed. Results: Exploration into the underlying mechanisms revealed that Ad5F35-SD-HA infection functioned by binding to the phospho-BCR-ABL Y177 site, which lead to a complex with Grb2. competitively disrupted the Grb2 SH2-phospho-BCR-ABL Y177 formation. The fusion protein SD-HA could reduce the activation of Ras and phosphorylation of MAPK (p-MAPK) and the expression level of p-ELK, inhibition of Ras-MAPK signaling pathway; SD-HA fusion protein could reduce p-Akt and Akt substrate p-GSK with inhibition of PI3K-Akt signaling pathway, thereby inhibiting the proliferation of K562 cells. Caspases-8-induced apoptosis signal could be activated by DED protein binding to DED domain of precursor caspases-8. Conclusions: The strategy of fusion protein SD-HA inhibiting-Y177 BCR-ABL and Grb2 binding could be used as a novel entry point for the treatment of chronic myeloid leukemia.
Adenoviridae ; Apoptosis ; Cell Proliferation ; Humans ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Oncogene Proteins, Fusion ; Phosphatidylinositol 3-Kinases

The study was aimed to explore whether there are leukemic characteristics in the bone marrow mesenchymal stem cells (BMMSC) from leukemic patients as compared with normal controls. The mesenchymal stem cells from bone marrow of normal volunteers and patients with APL and CML were isolated, then cultured and proliferated in vitro. The morphology, growth curve and cell surface markers of two different sources mesenchymal stem cells were investigated for detecting whether the bone marrow mesenchymal stem cells derived from leukemia patients have the specific abnormal fusion gene of leukemia cells through fluorescent in situ hybridization. The results indicated that there was no significant difference between the mesenchymal stem cells derived from different subjects, the bone marrow mesenchymal stem cells derived from leukemia patients did not have the clonal malignant fusion gene as seen in the leukemia cells. Taken altogether, mesenchymal stem cells derived from leukemia patients had no biological differences as compared with those from normal volunteers, and no malignant clonal abnormality was found. It is concluded that mesenchymal stem cells derived from leukemia patients as an alternative vehicle may be used for assistant of autologous hematopoietic stem cell transplantation or cell therapy and gene therapy.
Bone Marrow Cells ; cytology ; Cells, Cultured ; Fusion Proteins, bcr-abl ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; pathology ; Leukemia, Promyelocytic, Acute ; genetics ; pathology ; Mesenchymal Stromal Cells ; pathology ; Oncogene Proteins, Fusion ; genetics

To investigate the relationship between the single nucleotide polymorphism (SNPs) of the bcr and abl gene and chronic myelogeous leukemia (CML), the 9 sequence-tagged sites (STS) in bcr and abl gene were screened by DNA pooling and denaturing high performance liquid chromatography (dHPLC), and the results were varified by sequencing. The results showed that the polymorphism sites were detected in 4 out of the 9 STS fragments and there were 3 bases different from the reference sequence found in 3 fragments. In conclusion, the novel SNP in U07000 fragment shows significantly different frequencies between CML and controled people.
Chromatography, High Pressure Liquid ; methods ; Fusion Proteins, bcr-abl ; genetics ; Genes, abl ; genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Polymorphism, Single Nucleotide ; Proto-Oncogene Proteins c-bcr ; genetics ; Sequence Analysis, DNA ; Sequence Tagged Sites

This study was aimed to explore the single nucleotide polymorphism (SNPs) of breakpoint cluster region of bcr gene in Chinese people and the relationship between SNPs and chronic myelogenous leukemia (CML). A 3.12 kb region spanning from exon 13 to exon 15 in the bcr region were screened by DNA pooling and denaturing high performance liquid chromatography (dHPLC), and the results were verified by sequencing. The results indicated that 6 novel SNP sites and 2 bases different from reference sequence were confirmed in the region studied, and the frequency of 6 novel SNP sites in studied population was obtained, one SNP of which was found in exon 13 and caused a nonsynonymous mutation. The gene frequencies of novel SNPs had no significant difference between CML and control people. It is concluded that sequence polymorphisms in the major breakpoint cluster region of bcr gene are found, most of which are SNPs, No relationship can be confirmed between SNPs and CML disease.
Base Sequence ; Chromosome Breakage ; Fusion Proteins, bcr-abl ; genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Molecular Sequence Data ; Polymorphism, Single Nucleotide ; Proto-Oncogene Proteins c-bcr ; genetics

This study was aimed to investigate the effect of SU11248 on proliferation and apoptosis of leukemia cell line K562 in vitro and its mechanism. The inhibitory effect of 3.2 µg/ml SU11248 on K562 proliferation was tested by MTT assay. The ability of SU11248 to induce apoptosis of K562 cells was examined by TUNEL and DNA ladder. The expression of C-MYC, hTERT and BCR-ABL mRNA in K562 cells was detected by RT-PCR. The protein expression of Akt and p-Akt in K562 cells was detected by Western blot. The results showed that the proliferation of K562 cells was obviously inhibited by 3.2 µg/ml SU11248 in a time-dependent manner. SU11248 could induce K562 cells apoptosis in dose-and time-dependent manner. The mRNA expression of C-MYC, hTERT and BCR-ABL was reduced significantly by SU11248 in a time-dependent manner (P < 0.05). Western blot detection showed that the expression of p-Akt protein in K562 cells decreased in dose-and time-dependent manner after SU11248 treatment, but the expression of Akt was not significantly changed. It is concluded that SU11248 can inhibit the growth of K562 cells efficiently through inducing apoptosis, its mechanism may be closely relate with the expression down-regulation of C-MYC, hTERT, BCR-ABL and the inhibition of Akt phosphorylation.
Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Fusion Proteins, bcr-abl ; metabolism ; Humans ; Indoles ; pharmacology ; K562 Cells ; Proto-Oncogene Proteins c-akt ; metabolism ; Proto-Oncogene Proteins c-myc ; metabolism ; Pyrroles ; pharmacology ; RNA, Messenger ; genetics ; Telomerase ; metabolism

OBJECTIVETo establish a real-time quantitative reverse transcriptase polymerase chain reaction (RQ-RT-PCR) for quantitative detection of the common molecular markers that have affirmative clinical significance in the acute and chronic leukemia patients, and evaluate its significance in diagnosing leukemias and monitoring minimal residual disease (MRD).

METHODSPrimers and TaqMan probes were designed for detecting various fusion transcripts and normal abl gene was used as the internal control. The expression level of fusion transcripts in 202 newly diagnosed leukemias were determined.

RESULTSIn absolute quantity, expression level of the fusion transcripts in various leukemias was b3a2 (b2a2) 47614.63, e1a2 98847.53, AML1-ETO 300029.51, PML-RAR alpha 25506.28, respectively, while in relative quantity to abl, the levels were 1.05, 0.91, 5.33 and 0.55, respectively.

CONCLUSIONThe relative quantification of gene expression level by using RQ-RT-PCR to abl control gene is more accurate and direct viewing. Different levels of transcription of corresponding fusion genes are found in various subtypes of leukemias at diagnosis, among which the level of AML1-ETO was higher and PML-RAR alpha lower.

Biomarkers, Tumor ; genetics ; metabolism ; Core Binding Factor Alpha 2 Subunit ; genetics ; metabolism ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Humans ; Leukemia ; diagnosis ; genetics ; metabolism ; Oncogene Proteins, Fusion ; genetics ; metabolism ; RUNX1 Translocation Partner 1 Protein ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription, Genetic

OBJECTIVETo explore the application of combined detection of fusion gene and BIOMED-2 standardized immunoglobulin (Ig) gene rearrangement system in diagnosis and treatment of children with acute lymphoblastic leukemia (ALL).

METHODSMultiplex-PCR amplifications and RQ-PCR of RNA/DNA were performed using ALL fusion gene detection kit and BIOMED-2 primer. The Ig gene rearrangements were analyzed by using PCR fragment analysis system.

RESULTSOut of 251 children with B-ALL, 77 cases were TEL-AML1(+) , 28 cases were E2A-PBX1(+) , 10 cases were MLL-AF4(+) , 11 cases were BCR-ABL(+) , the total positive rate was 50.2%, 82.5% showed IgH VH-JH rearrangement, 53.4% showed IgK rearrangement. The positive rate of combined detection of fusion gene and gene rearrangement was 99%. E2A-PBX1(+) and MLL-AF4(+) with IgK(+) gene rearrangement group was compared with negative control group, the difference was statistically significant (P < 0.001 or P = 0.005); 105 ALL fusion gene positive cases had been detected by fluorescence in situ hybridization (FISH) simultaneously, the accordance rate of fusion gene and FISH was more than 94%.

CONCLUSIONThe combined detection of ALL fusion gene and BIOMED-2 standardized clonality analysis system can improve the positive detected rate of B-ALL dramatically, and make the grouping of disease prognosis more accurately; this combined detection is a more faster and sensitive method than FISH.

Child ; Core Binding Factor Alpha 2 Subunit ; genetics ; DNA Primers ; Fusion Proteins, bcr-abl ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Multiplex Polymerase Chain Reaction ; Oncogene Proteins, Fusion ; genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics ; V(D)J Recombination

OBJECTIVE: To analyze 43 leukemia genes in children with acute lymphoblastic leukemia (ALL) in Yunnan province, and provide the basis for the diagnosis and treatment of children with ALL in this area. METHODS: The clinical data of 428 children with newly diagnosed ALL in Yunnan area from January 2015 to December 2020 were retrospectively analyzed. Multiple nested PCR technology was used to detect 43 common leukemia genes. RESULTS: Among the 428 children with ALL, 159 were positive for leukemia genes, with a positive rate of 37.15% (159/428), and a total of 15 leukemia genes were detected. Among the 159 leukemia gene-positive children, ETV6-RUNX1⁺ accounted for 25.79% (41/159), followed by E2A-PBX1⁺ and BCR-ABL⁺, accounting for 24.53% (39/159) and 23.27% (37/159) respectively. MLL⁺ accounted for 6.29% (10/159), WT1⁺ accounted for 4.40% (7/159), IKZF1 gene deletion and CRLF2⁺ accounted for 3.77% (6/159) respectively. The positive rate of MLL (46.15%) was the highest in ＜1-year old group, the positive rate of ETV6-RUNX1 (10.56%) was the highest in 1-10-year old group, and BCR-ABL⁺ rate (23.65%) was the highest in >10-year old group. The distribution of leukemia genes in different age groups was statistically significant (P <0.05). CONCLUSION The most common fusion gene of children with ALL in Yunnan is ETV6-RUNX1, followed by E2A-PBX1 and BCR-ABL.
Child ; Humans ; Infant ; Child, Preschool ; Oncogene Proteins, Fusion/genetics* ; Fusion Proteins, bcr-abl/genetics* ; Core Binding Factor Alpha 2 Subunit/genetics* ; Retrospective Studies ; China ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/therapy* ; Genotype