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1.
Journal of Experimental Hematology ; (6): 1436-1449, 2021.
Article in Chinese | WPRIM | ID: wpr-922277

ABSTRACT

OBJECTIVE@#LncRNA ITGB2-AS1 has been found to play important roles in the occurrence and development of human solid tumors. However, its role in hematological diseases, especially acute myeloid leukemia (AML), remains unclear. The aim of this study was to identify the expression pattern of ITGB2-AS1 in AML patients and to further explore its clinical significance.@*METHODS@#ITGB2-AS1 expression was analyzed in public datasets (including TCGA and GSE63270) and further validated in a cohort of 109 AML patients by real-time quantitative PCR (RT-qPCR).@*RESULTS@#The level of ITGB2-AS1 was up-regulated among two independent cohorts (TCGA, P<0.05; GSE63270, P<0.05), which was confirmed by the data from 109 AML patients enrolled in this study (P<0.05). Clinically, high ITGB2-AS1 expression was associated with older age (P=0.023) and lower complete remission (CR) rate (P=0.005). Multivariate analysis identified that high ITGB2-AS1 expression was an independent prognostic factor not only for CR rate (P=0.027) but also for overall survival (OS) time (P=0.011), and ITGB2-AS1 was positively correlated with ITGB2 expression in both TCGA (r=0.74, P<0.001) and clinical data detected in this study (r=0.881, P<0.001). High ITGB2 expression was also associated with older age (P=0.02) and lower CR rate (P=0.020). Moreover, high ITGB2 expression predicted worse OS (P=0.028).@*CONCLUSION@#ITGB2-AS1 is overexpressed in AML and predicts poor prognosis in AML patients.


Subject(s)
Aged , Humans , Leukemia, Myeloid, Acute/genetics , Prognosis , RNA, Long Noncoding/genetics
2.
Article in Chinese | WPRIM | ID: wpr-272504

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the methylation status of miR-378 promoter in chronic myeloid leukemia (CML) and to analyze its clinical significance.</p><p><b>METHODS</b>The unmethylation level of miR-378 gene promoter in bone marrow mononuclear cells of 25 healthy donors and 53 patients with CML was detected by using real-time quantitative methylation-specific PCR (RQ-MSP).</p><p><b>RESULTS</b>The hypomethylation of miR-378 gene promoter was found in 17/53 (32.1%) patients, but only in 1/25 (4.0%) of controls. The difference between the two groups was very statistically significant (P < 0.01). The frequency of miR-378 unmethylation in CML patients at chronic phase (CP), accelerated phase (AP) and blastic phase (BP) was 35.0% (14/40), 40.0% (2/5), and 12.5% (1/8), respectively. However, there were no significant differences in the unmethylation level of miR-378 among CML patients at different sexes, stages and karyotypes. No significant differences could be observed in age, white blood cell counts, platelet count, hemoglobin level and BCR/ABL1 transcript level (P > 0.05). CONCLUDSION: The miR-378 hypomethylation is a common molecular event in CML, especially at chronic or accelerated phases.</p>


Subject(s)
Bone Marrow Cells , Metabolism , Case-Control Studies , DNA Methylation , Humans , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Genetics , MicroRNAs , Metabolism , Promoter Regions, Genetic
3.
Journal of Experimental Hematology ; (6): 1111-1115, 2013.
Article in English | WPRIM | ID: wpr-283971

ABSTRACT

This study was aimed to investigate the expression pattern of gene PDLIM4 (PDZ and LIM domain 4) and analyze its clinical correlation with the patients suffered from acute myeloid leukemia (AML). The expression pattern of PDLIM4 in AML was detected by using EvaGreen real-time quantitative PCR (RQ-PCR). The results showed that the PDLIM4 transcript significantly decreased in 94 AML patients, compared with 21 controls (P < 0.01). The decrease of PDLIM4 transcript was found in 42 (45%) AML patients. PDLIM4 low-expression occurred among the subtypes of M1/M2/M3 more frequently than that in M4/M5/M6 (56% vs 20%, P < 0.01). AML patients with PDLIM4 low-expression had an overall survival (OS) higher than that in AML patients without PDLIM4 low-expression (P < 0.05). Analysis with receiver operating characteristic curve (ROC) displayed that PDLIM4 expression possesses the diagnostic value to differentiate AML from controls, with ROC curve area of 0.865 (95% CI: 0.801-0.930). It is concluded that reduced PDLIM4 expression is a common and favorable event for the good prognosis in AML, and can be used as a potential diagnostic biomarker of cancer.


Subject(s)
Adolescent , Adult , Aged , Biomarkers, Tumor , Metabolism , Case-Control Studies , DNA-Binding Proteins , Genetics , Metabolism , Female , Humans , LIM Domain Proteins , Genetics , Metabolism , Leukemia, Myeloid, Acute , Diagnosis , Genetics , Metabolism , Male , Middle Aged , Prognosis , Young Adult
4.
Article in Chinese | WPRIM | ID: wpr-325220

ABSTRACT

The purpose of this study was to detect the expression of RAGE-1 transcript in the bone marrow mononuclear cells (BMMNC) from patients with acute myeloid leukemia (AML) and to investigate the relationship of RAGE-1 expression level with clinical variables. The expression level of RAGE-1 gene in BMMNC from 94 newly diagnosed AML patients was measured using RQ-PCR. The relationship between RAGE-1 expression level and clinical parameters (age, sex, blood cell counts, diagnosis and prognosis) was investigated, and the levels of RAGE-1 expression were compared in patients before and after treatment. The results showed that overexpression of RAGE-1 transcript was found in 28% (26/94) AML patients (1.34 - 16.34, median 3.07). No significant difference was observed in sex, age, blood parameters and FAB subtypes between the groups with and without RAGE-1 overexpression. There was also no significant difference in the frequency of RAGE-1 overexpression among different cytogenetic risk groups and among the patients with different types of karyotypes. The level of RAGE-1 transcript significantly decreased in those patients obtained complete remission after treatment. The overall survival of AML patients with RAGE-1 overexpression was similar as that in those without RAGE-1 overexpression. It is concluded that RAGE-1 overexpression is a common event in AML, but has no impact on the prognosis of patients.


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Cell Line, Tumor , Female , Gene Expression , Humans , Karyotyping , Leukemia, Myeloid, Acute , Genetics , Male , Middle Aged , Prognosis , Young Adult
5.
Journal of Experimental Hematology ; (6): 1171-1175, 2011.
Article in English | WPRIM | ID: wpr-261907

ABSTRACT

The aim of this study was to investigate the expression status of the helicase antigen (HAGE) transcript and its clinical significance in patients with acute myeloid leukemia (AML) and chronic myeloid leukemia (CML). The expression of HAGE cDNA in bone marrow mononuclear cells from AML and CML patients was detected by using real-time quantitative PCR. The results indicated that overexpression of HAGE transcript (117.12% - 9842.70%, median 434.96%) was detected in 14.8% (11/74) AML patients. AML patients with HAGE cDNA expression were significantly older than those HAGE-negative patients (median 67 and 45 years, respectively, p = 0.001). HAGE cDNA expression was more frequently present among the patients with acute monoblastic leukemia (M(4) and M(5), 7 of 20, 35.0%), compared to the patients with acute non-monoblastic leukemia (M(1), M(2), M(3) and M(6), 4 of 54, 7.4%) (p = 0.007). 28.6% (8/28) cases with normal karyotypes showed HAGE cDNA overexpression, significantly higher than 7.5% (3 of 40) in those with chromosomal abnormalities (p = 0.041). Overexpression of HAGE transcript was found in 9 (34.6%) CML cases and more frequently observed at accelerated phase and blast crisis (4/4, 100%) than that at chronic phase (5/22, 22.7%) (p = 0.008). It is concluded that HAGE cDNA expression is relevant to specific subtypes of AML and to the progression of CML.


Subject(s)
Blast Crisis , Bone Marrow Cells , Metabolism , DEAD-box RNA Helicases , Genetics , Metabolism , DNA, Complementary , Gene Expression , Humans , Karyotype , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Genetics , Metabolism , Leukemia, Myeloid, Acute , Genetics , Metabolism , Neoplasm Proteins , Genetics , Metabolism , RNA, Messenger , Genetics
6.
Chinese Journal of Hematology ; (12): 21-24, 2011.
Article in Chinese | WPRIM | ID: wpr-252021

ABSTRACT

<p><b>OBJECTIVE</b>To detect the common mutations (D816V and N822K) of c-kit gene in acute myeloid leukemia (AML) using high-resolution melting analysis (HRM).</p><p><b>METHODS</b>HRM analysis was established to screen c-kit mutations in PCR products of c-kit exon 17 in 21 AML patients with t(8;21). PCR products were sequenced to confirm the mutation.</p><p><b>RESULTS</b>HRM analysis identified an aberrant melting curve in 6 cases (28.6%), which were confirmed by direct DNA sequencing as one D816V mutation and five N822K mutation.</p><p><b>CONCLUSION</b>HRM analysis is a convenient, rapid, specific and high-throughput technique for scanning c-kit gene mutation in AML.</p>


Subject(s)
Adolescent , Adult , Aged , Child , DNA Mutational Analysis , Methods , Exons , Female , Humans , Leukemia, Myeloid, Acute , Genetics , Male , Middle Aged , Mutation , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Genetics , Proto-Oncogene Proteins c-kit , Genetics , Young Adult
7.
Article in Chinese | WPRIM | ID: wpr-349023

ABSTRACT

<p><b>OBJECTIVE</b>To analyze the expression level and clinical significance of the preferentially expressed antigen of melanoma (PRAME) transcripts in patients with acute myeloid leukemia (AML).</p><p><b>METHODS</b>Real-time quantitative polymerase chain reaction with EvaGreen dye was established to detect the expression level of PRAME transcripts in the bone marrow mononuclear cells of 56 AML cases and 20 controls. The clinical association of PRAME transcripts was analyzed.</p><p><b>RESULTS</b>The PRAME transcripts were 0-1.46% (median 0.18%) and 0-21 618.09% (median 9.79%) in controls and AML cases, respectively (P< 0.01). Among the FAB subtypes, those with M1, M2, M3 and M4 had significantly higher level of PRAME transcripts than controls. However, those with M5 had similar level of PRAME transcripts as controls. There was a significantly negative correlation between the PRAME transcripts and cytogenetic risk groups (r= -0.438, P= 0.001). Cases in low risk had significantly higher level of PRAME transcripts than those in intermediate and high risk. Among cases with AML-M2, those with t(8;21) had significantly higher level of PRAME transcripts (135.06% -21 618.09%, median 2201.88%) than those without t(8;21)(0.14% -1696.30%, median 17.97%)(P= 0.002). In a patient with sequential samples, PRAME transcripts significantly decreased after induction therapy and significantly increased after relapse.</p><p><b>CONCLUSION</b>The PRAME transcript was highly expressed in AML patients and was a favorable marker of prognosis. Quantification of PRAME transcript can be used in monitoring disease status of AML.</p>


Subject(s)
Antigens, Neoplasm , Genetics , Case-Control Studies , Cytogenetic Analysis , Female , Genetic Predisposition to Disease , Humans , Leukemia, Myeloid, Acute , Genetics , Therapeutics , Male , Polymerase Chain Reaction , RNA, Messenger , Genetics , Recurrence
8.
Article in Chinese | WPRIM | ID: wpr-348994

ABSTRACT

<p><b>OBJECTIVE</b>To quantify the expression level of GRAF gene in acute myeloid leukemia (AML) and analyze its clinical significance.</p><p><b>METHODS</b>The EvaGreen real-time quantitative polymerase chain reaction (RQ-PCR) assay was established and performed to measure the GRAF gene transcripts in 71 cases with AML and 21 with nonmalignant hematological diseases. The clinical correlation of GRAF expression was analyzed.</p><p><b>RESULTS</b>The established EvaGreen RQ-PCR assay had good specificity, reproducibility and sensitivity. The GRAF expression level was significantly lower in AML (0.01%-169.75%, median 3.82%) than that in controls (14.49%-126.85%, median 56.04%) (P<0.05). There was no correlation between the level of GRAF transcript and the sex, age, hematologic parameters, FAB subtypes and karyotypic groups (P>0.05).</p><p><b>CONCLUSION</b>The GRAF gene was down-regulated in AML, which might play a role in the leukemogenesis.</p>


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Female , GTPase-Activating Proteins , Genetics , Humans , Leukemia, Myeloid, Acute , Genetics , Male , Middle Aged , Polymerase Chain Reaction , Methods , Young Adult
9.
Article in Chinese | WPRIM | ID: wpr-237637

ABSTRACT

This study was purposed to analyze the expression level of preferentially expressed antigen of melanoma (prame) transcript in the patients with chronic myeloid leukemia (CML) and explore its clinical significance. Real-time quantitative PCR (RQ-PCR) assay was used to detect the level of prame gene transcript in the bone marrow samples from 30 patients with CML and 15 patients with iron deficiency anemia (IDA). The results showed that CML patients had significantly higher prame mRNA level (0% - 772.25%, median 8.28%) than IDA cases (0% - 1.46%, median 0.19%) (p < 0.001). The level of prame gene transcript was significantly correlated with that of bcr-abl fusion gene transcript (r = 0.708, p < 0.001) in CML patients. Furthermore, 6 patients in blastic crisis (BC) and accelerated phase (AP) had significantly higher prame gene transcript than that of 24 cases in chronic phase (CP) (p = 0.007). In 2 CML patients with sequential samples, prame gene transcript increased in AP and BC, compared with in CP, and was consistent with the altering tendency of bcr-abl transcript. It is concluded that the level of prame gene transcript increases in CML which associates with the progression of the disease, prame gene transcript level can be used for monitoring the disease state.


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Antigens, Neoplasm , Genetics , Asians , Genetics , Child , Female , Humans , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Genetics , Male , Middle Aged , Reverse Transcriptase Polymerase Chain Reaction , Methods , Transcription, Genetic , Young Adult
10.
Journal of Experimental Hematology ; (6): 1390-1394, 2010.
Article in Chinese | WPRIM | ID: wpr-332353

ABSTRACT

This study was purposed to analyze the methylation status of death-associated protein kinase (dapk) gene promoter in Chinese patients with acute myeloid leukemia (AML) and its relationship with clinical features. The methylation-specific PCR (MSP) technique was used to detect dapk promoter methylation in bone marrow samples from 112 cases of AML. The results indicated that gene dapk promoter hypermethylation was detected in 82 cases (73.2%), but not in 13 control group. There was no correlation of dapk gene hypermethylation with sex, age, WBC counts, platelet counts, hematologic parameters, chromosomal abnormalities and different subtypes of AML patients. It is concluded that dapk gene hypermethylation may be a common molecular event in AML.


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Apoptosis Regulatory Proteins , Genetics , Calcium-Calmodulin-Dependent Protein Kinases , Genetics , Child , Child, Preschool , DNA Methylation , Death-Associated Protein Kinases , Female , Humans , Leukemia, Myeloid, Acute , Genetics , Male , Middle Aged , Promoter Regions, Genetic , Young Adult
11.
Article in Chinese | WPRIM | ID: wpr-229825

ABSTRACT

<p><b>OBJECTIVE</b>To study the methylation status of fragile histidine triad (FHIT) gene promoter in patients with myelodysplastic syndrome (MDS) and its clinical relevance.</p><p><b>METHODS</b>Methylation-specific PCR (MSP) was used to detect FHIT promoter methylation in bone marrow samples from 54 MDS cases.</p><p><b>RESULTS</b>Hypermethylation of FHIT promoter was detected in 26 cases (48.1%). Association was not found between FHIT gene hypermethylation and sex, hematologic parameters and chromosomal abnormalities of MDS patients, but found between FHIT gene hypermethylation and age of the MDS cases. Although significant difference was not observed in the frequencies of FHIT gene hypermethylation among patients with refractory anemia/refractory anemia with ringed sideroblasts (RA/RAS) (1/6, 16.7%), refractory anemia/refractory anemia with ringed sideroblasts (RCMD) and refractory cytopenia with multilineage dysplasia with ringed blasts (RCMD-RS) (6/19, 31.6%), refractory anemia with excess blasts-1 (RAEB-1) (7/11, 63.6%), refractory anemia with excess blasts-2 (RAEB-2) (4/7, 57.1%) and refractory anemia with excess blasts in transformation/acute myeloid leukemia (RAEBt/AML) (8/11, 72.7%)(chi-square=8.417, P=0.077), it was observed in patients in early stages (RA/RAS and RCMD) (7/25, 28.0%), advanced stages (RAEB-1 and RAEB-2)(11/18, 61.1%) and RAEBt/AML (8/11, 72.7%) (chi-square=7.938, P=0.019). Furthermore, there was a positive correlation between the frequency of FHIT gene hypermethylation and different IPSS groups (chi-square=10.110, P=0.018).</p><p><b>CONCLUSION</b>FHIT gene hypermethylation might be one of the molecular events involved in the disease progression of MDS.</p>


Subject(s)
Acid Anhydride Hydrolases , Genetics , Adolescent , Adult , Age Factors , Aged , Aged, 80 and over , Base Sequence , DNA Methylation , Female , Humans , Male , Middle Aged , Molecular Sequence Data , Myelodysplastic Syndromes , Classification , Genetics , Pathology , Neoplasm Proteins , Genetics , Polymerase Chain Reaction , Promoter Regions, Genetic , Genetics
12.
Chinese Journal of Hematology ; (12): 753-756, 2008.
Article in Chinese | WPRIM | ID: wpr-239961

ABSTRACT

<p><b>OBJECTIVE</b>To establish the real time quantitative PCR (RQ-PCR) assay according to 'Europe Against Cancer' (EAC) program and analyze the results of detection and quantification of different PML-RAR alpha transcript isoforms in patients with acute promyelocytic leukemia (APL).</p><p><b>METHODS</b>Three RQ-PCR systems were performed to detect the most frequent PML-RAR alpha transcripts (L-form, S-form and V-form) in 30 APL patients and the RQ-PCR end-point products were identified by electrophoresis.</p><p><b>RESULTS</b>S-form RQ-PCR system could amplify positive signals of three isoforms in all of 30 cases, and V-form RQ-PCR system could do so in both L-form and V-form positive cases, however, L-form RQ-PCR system could only do so in L-form-positive cases. Electrophoresis and sequencing of end-point products amplified by S-form RQ-PCR system revealed three bands in each of L-form (621 bp, 477 bp and 218 bp) and V-form (567 bp, 423 bp and 218 bp) positive patients samples.</p><p><b>CONCLUSIONS</b>RQ-PCR, sensitive and reliable, can be used for monitoring the minimal residual disease in APL patients, however, its results should be interpreted carefully if it is used for detection of PML-RAR alpha fusion transcripts prospectively.</p>


Subject(s)
Adolescent , Adult , Child , Female , Humans , Leukemia, Promyelocytic, Acute , Diagnosis , Genetics , Male , Middle Aged , Neoplasm, Residual , Diagnosis , Genetics , Oncogene Proteins, Fusion , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Methods , Sensitivity and Specificity , Young Adult
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