The study was purposed to investigate the significance of id4 gene promoter methylation in monitoring AML patients with complete remission (CR). Methylation specific-PCR (MS-PCR) were used to detect the status of promoter methylation of id4 gene in bone marrow samples from AML patients with CR who had accepted induction with DA or IA and 4 to 5 consolidation chemotherapy with Ara-C. The results showed that in the all 32 patients, 15 were found to show id4 promoter methylation and 7 out of the 15 were found relapsed or tendency to relapse in the following-up period. While all the 17 patients with id4 unmethylation were still in their CR status in the same period. In conclusion, id4 promoter methylation might be a predictor for relapse of AML patients with CR in certain degree.
DNA Methylation ; Humans ; Inhibitor of Differentiation Proteins ; genetics ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Neoplasm, Residual ; diagnosis ; genetics ; Promoter Regions, Genetic ; genetics

The study was aimed to investigate the expression of preferentially expressed antigen of melanoma (PRAME) gene in adult acute leukemia and its clinical significance. The expression of the PRAME gene of bone marrow was measured by reverse transcriptase polymerase chain reaction (RT-PCR) in 73 adult newly diagnosed acute leukemia patients, 3 relapsed patients, 7 patients with idiopathic thrombocytopenic purpura (ITP) and 8 healthy donors, as well as two AL cell-lines (K562 and U937). The results indicated that PRAME mRNA was expressed in 42.9% AML patients (n=24) and 20% ALL patients (n=4), also in two leukemia cell-lines K562 and U937, but not in eight health donors and seven ITP patients. PRAME expression not correlated to the white blood count, hemoglobin level, platelet count and the percentage of blasts at diagnosis, yet independent of age, sex, and FAB type. PRAME mRNA expression in complete remission group seems much higher than those in partial complete remission group and death group. The increased levels of expression could be found prior to the relapse in one patient being regularly monitored. PRAME gene was overexpressed in adult acute leukemia patients and leukemia cell-lines. It is concluded that the expression of PRAME is an indicator of favorable prognosis and can be a useful tool for monitoring minimal residual disease (MRD) in adult acute leukemia. Differential expression between adult acute leukemia patients and healthy volunteers suggests that the immunogenic antigens PRAME are potential candidates for immunotherapy in adult acute leukemia.
Acute Disease ; Adolescent ; Adult ; Aged ; Antigens, Neoplasm ; metabolism ; Female ; Humans ; Leukemia ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Prognosis ; RNA, Messenger ; metabolism ; Young Adult

OBJECTIVE: To investigate the significance of human thioredoxin-2 (TRX-2) in monitoring minimal residual disease (MRD) in acute leukemia (AL). METHODS: We used real-time quantitative PCR to serially quantitize TRX-2 expression levels in the bone marrow of AL patients at diagnosis (n=68), at complete hematologic remission (CHR, n=57) and at relapse (n=25). Another 25 normal donors served as normal controls. The upper limit of the bone marrow at 91 was regarded as the reference. TRX-2 expression level at CHR with <5% blast cells in the bone marrow of relapse patients was analyzed and compared with MRD by flow cytometry. RESULTS: The TRX-2 levels between the CHR patients and newly diagnosed patients, and between the CHR patients and the relapse patients had significant difference. TRX-2 expression level of 21(21/25) relapse patients at CHR with <5% blast cells in the bone marrow was higher than the reference (>91). TRX-2 level was correlated to the expression level of MRD. CONCLUSION TRX-2 may be the marker for AL and used in MRD monitoring.
Case-Control Studies ; Female ; Humans ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; metabolism ; Male ; Neoplasm, Residual ; diagnosis ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; Thioredoxins ; genetics ; 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

The purpose of this study was to detect the serum PTK7 level of patients with acute lymphocytic leukemia, and to reveal its clinical value for diagnosis of diseases. A total of 136 patients diagnosed as acute lymphocytic leukemia from May 2012 to April 2014 in our hospital were enroled in this study and were divided into the L1 group (n = 42), L2 (n = 45) and L3 group (n = 49) according cytomorphology, and 48 normal children were selected as control group. Fluorescence quantitative PCR was used to detect mRNA level of PTK7 in peripheral blood mononuclear cells, and Western blot was used to detect PTK7 protein expression. The results showed that the PTK7 mRNA level in L1 group was significantly higher than that in normal group (P = 0.000) . The PTK7 mRNA level in L2 group was significantly higher than that in the L1 group (P = 0.000). The PTK7 mRNA level in L3 group and L2 group had not significantly different between each other (P = 0.123). Serum PTK7 protein level in L1 group was very significantly higher than that in normal group (P = 0.000) . The serum PTK7 protein level in L2 group were very significantly higher than that in the L1 group (P = 0.003) and serum PTK7 protein level in L3 and L2 group had no significance difference (P = 0.312) . It is concluded that the expression level of serum PTK7 protein has a potential clinical value for the diagnosis of acute lymphocytic leukemia, but without specificity for ALL subsets.
Cell Adhesion Molecules ; blood ; genetics ; Humans ; Leukocytes, Mononuclear ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; diagnosis ; genetics ; RNA, Messenger ; biosynthesis ; blood ; genetics ; Receptor Protein-Tyrosine Kinases ; blood ; genetics

OBJECTIVETo investigate the feasibility of detecting cyclin D1 mRNA in paraffin-embedded tissues by reverse transcriptase polymerase chain reaction (RT-PCR) and competitive RT-PCR and its diagnostic and differential diagnostic significance for mantle cell lymphoma (MCL).

METHODSParaffin-embedded samples of 36 cases of MCL, 71 cases of other small B-cell lymphomas and 20 cases of lymphoid reactive hyperplasia as control group were retrieved from archival materials. Cyclin D1 protein and its mRNA was detected by EnVision and RT-PCR and competitive RT-PCR in all samples. House-keeping gene PGK was choosen as internal control.

RESULTS(1) Cyclin D1 protein was expressed in 27 of the 38 MCL (71.1%). No cyclin D1 expression was found in the control group. (2) PGK was detected in 103 of the 116 cases (88.8%) and also detected in 34 of 36 MCL cases (94.7%). (3) cyclin D1 mRNA was detected in 34 nodal mantle cell lymphoma cases by RT-PCR in paraffin-embedded tissues. The positive rate of cyclin D1 mRNA was 94.4% in mantle cell lymphomas after exclusion of the 2 cases which were negative for both cyclin D1 mRNA and PGK. cyclin D1 mRNA was not detected in other nodal small B-cell lymphomas or lymphoid reactive hyperplasia, except 1 case of B-SLL. Sequencing analysis showed that sequences were identical to cyclin D1. (4) Cyclin D1 mRNA overexpression was detected in 27 cases of nodal mantle cell lymphoma by competitive RT-PCR in paraffin-embedded tissues. The positive rate of cyclin D1 mRNA overexpression was 75.0% in mantle cell lymphomas after exclusion of 2 cases which were negative for both cyclin D1 mRNA and PGK. cyclin D1 mRNA overexpression was not detected in other nodal small B-cell lymphomas or lymphoid reactive hyperplasia.

CONCLUSIONRT-PCR and competitive RT-PCR detection of cyclin D1 mRNA overexpression could be used for the diagnosis and differential diagnosis of mantle cell lymphoma in paraffin-embedded blocks.

Cyclin D1 ; biosynthesis ; genetics ; Diagnosis, Differential ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; metabolism ; pathology ; Lymphoma, Follicular ; genetics ; metabolism ; Lymphoma, Mantle-Cell ; genetics ; metabolism ; pathology ; Paraffin Embedding ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods

No abstract available.
Aged ; B-Lymphocytes/*cytology/metabolism ; Child ; *Chromosomes, Human, Pair 11 ; *Chromosomes, Human, Pair 9 ; Female ; Humans ; Immunophenotyping ; Infant ; Karyotyping ; Male ; Myeloid-Lymphoid Leukemia Protein/genetics/metabolism ; Oncogene Proteins, Fusion/genetics/metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/*diagnosis ; Translocation, Genetic

Diagnosis, Differential ; Female ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Leukemia, Myeloid, Chronic-Phase ; genetics ; Lymphoma ; classification ; diagnosis ; pathology ; Male ; Neoplasms ; genetics ; metabolism ; pathology ; Oncogene Proteins ; metabolism ; Prognosis ; Prostatic Neoplasms ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Translocation, Genetic ; Tumor Suppressor Proteins ; metabolism

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.
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

This study was aimed to investigate the chimerism and fusion gene expression in patients with CML after allo-HSCT, to analyse engraftment and minimal residual disease by using STR-PCR combined with RT-PCR qualitative and quantitative assays, and to evaluate their clinical value for predicting disease relapse. 4 relapsed patients with CML after allo-HSCT were dynamically investigated. Qualitative analysis of donor chimerism was performed by multiplex PCR amplification of STR markers and capillary electrophoresis with fluorescence detection, qualitative detection of bcr/abl transcripts was performed by RT-PCR. The results showed that the 100% donor chimerism appeared in 4 patients on day 28 after transplantation and bcr/abl expression was negative, but the 4 patients were in status of unstable mixed chimerism (DC: 0% - 80.4%) at the different time points during the following up with bcr/abl gene positive. 2 patients of them were continuously mixed chimerism after relapse of CML, the other 2 changed from MC to CC by intervention of clinical treatment. Decreasing values of donor chimerism were detected prior to the occurrence of graft rejection and CML relapse, and bcr/abl gene expression was positive. It is concluded that the results of STR-PCR in the range of its sensitivity fully correspond with bcr/abl tests in patients. The combination of STR-PCR with RT-PCR will provide a highly sensitive and valuable tool for evaluating engraftment, graft rejection, and relapse and predicting GVHD. Furthermore, it can provide a basis for early intervention of clinical treatment, and can identify these high risk patients with molecular or cytogenetic relapse after allo-HSCT.
Fusion Proteins, bcr-abl ; genetics ; metabolism ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; therapy ; Neoplasm Recurrence, Local ; genetics ; Neoplasm, Residual ; diagnosis ; genetics ; RNA, Messenger ; genetics ; metabolism ; Transplantation Chimera ; Transplantation, Homologous