In recent years, standardized treatment based on the risk stratification has been applied to clinical diagnosis and treatment of leukemia, which significantly improves the remission rate of ALL. However, relapse after remission remains an important challenge for long term efficacy. Chromosomal karyotype analysis is often used clinically to study the genetic features of ALL. As leukemia-specific markers, the cytogenetic and molecular genetic abnormalities can be used to evaluate prognosis and make an effective and optimal therapy. Furthermore, they are also used to track minimal residual disease. Therefore, the cytogenetic and molecular genetic abnormalities may become a monitor and a new target for the treatment of leukemia. This review briefly introduces the structure and physiological function of B-ALL associated cytogenetic and molecular genetic abnormalities, focusing on their prognostic effect on B-ALL.
Cytogenetic Analysis ; Humans ; Karyotyping ; Neoplasm, Residual ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics ; Prognosis

OBJECTIVETo set up the technical system of multiplex fluorescence in situ hybridization M-FISH and to explore its application in detection of the complex chromosome abnormalities in leukemia.

METHODSThe complex chromosome abnormalities of two leukemia patients were analyzed by the combination use of classical cytogenetics, chromosome painting (CP), FISH and M-FISH.

RESULTSIn a case of acute lymphoblastic leukemia-L2, the complex karyotype: 46,XY,der(2)t(2;9),der(9)t(9;12;22) was identified by M-FISH, which was detected as 46,XY,der(9)t(9;12) by classical cytogenetics; In a case of acute monocytic leukemia-M5, the complex chromosome abnormalities: 46,XY,der(2)t(2;17), der(10)t(10;11;17), der(11)t(11;?) was revealed by M-FISH, which was confirmed by CP and FISH, and mixed lineage leukemia (MLL) gene was also found involved in this complex chromosome translocation.

CONCLUSIONM-FISH was proved to be a powerful tool to examine the complicated karyotypes and hopefully to elucidate nearly all chromosomal aberrations in leukemia and other cancers.

Chromosome Aberrations ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics

INTRODUCTIONThe analysis of immunophenotype of the leukaemic cells has been of great importance for the diagnosis, classification and prognosis of acute lymphoblastic leukaemia (ALL).

MATERIALS AND METHODSOne hundred and thirteen Chinese patients with ALL were immunophenotyped by fl ow cytometry and 74 cases were also subjected to karyotype analysis by G-banding technology.

RESULTSOf the 113 Chinese ALL patients, 14.2% were identified as T-ALL and 85.8% as B-ALL. Myeloid antigen (MyAg) expression was documented in 34.9% of the cases analysed and CD13 was most commonly expressed MyAg in ALL patients (23.6%). MyAg positivity was higher in adult with ALL (47.6%) than in children with ALL (26.6%). Abnormal karyotypes were detected in 39 out of 74 (52.7%) cases. The clinical and biological characteristics of ALL patients between MyAg+ and MyAg- groups showed that increased white blood count (WBC) (>50 x 109/L), higher CD34 positivity and higher percentage of adult patients were found to be correlated with MyAg+ ALL.

CONCLUSIONOur results indicate that the immunophenotype did have relevance to the abnormal cytogenetic changes and clinical features in ALL. Flow cytometry immunophenotype has become the most important method for diagnosis and typing of ALL.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Cytogenetic Analysis ; Diploidy ; Female ; Humans ; Immunophenotyping ; Infant ; Male ; Middle Aged ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ; classification ; diagnosis ; genetics ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; classification ; diagnosis ; genetics ; Translocation, Genetic ; Young Adult

OBJECTIVETo explore detection of immunoglobulin heavy chain gene (IgH) rearrangement by real-time quantitative polymerase chain reaction (RQ-PCR) for minimal residual disease (MRD) monitoring in adult B-lineage acute lymphoblastic leukemia (B-ALL) patients.

METHODSDNA samples of fifteen newly diagnosed adult B-ALL patients were collected. The IgH gene rearrangements were detected by PCR followed by sequencing and subsequent blasting for monoclonal PCR products. Allele-specific oligonucleotides (ASO) were designed based on the sequence of junction regions, using PRIMER 5.0 software. MRD targets were detected in 115 bone marrow samples by RQ-PCR, in which ASO upstream primers in combination with the consensus JH probes and downstream primers were used. Transcripts copies of bcr-abl fusion gene were also measured in 7 Ph(+) ALL cases.

RESULTSThe detection sensitivity of ASO-PCR varied between 10(-3) and 10(-5) leukemia cells in 15 adult ALL patients. The background and nonspecific amplification was detectable at a low level. Quantification monitoring MRD showed that high-risk adult ALL patients in complete remission (CR) had a higher MRD level than those of standard-risk. Patients with MRD > 10(-3) had a higher relapse rate and a shorter survival time. Besids, the dynamic curves of the quantified level of respective IgH rearrangement were consistent with the expression levels of bcr-abl fusion genes in seven Ph(+) patients during follow-up.

CONCLUSIONSThe individual quantification of IgH rearrangement by RQ-PCR using ASO primers was a sensitive, specific and reliable method for accurate evaluation of malignant clones. These data indicates a close correlation between the level of rearranged IgH and the treatment response and prognosis in adult ALL patients. It may be a helpful method for monitoring MRD in clinical trials.

Adult ; DNA Primers ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulins ; therapeutic use ; Neoplasm, Residual ; diagnosis ; Polymerase Chain Reaction ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics

OBJECTIVETo evaluate the three-probe fluorescence in situ hybridization (FISH) panel in the diagnosis of pediatric B cell acute lymphoblastic leukemia (B-ALL).

METHODSThree-probe (TELAML1, BCR-ABL and MLL) FISH and conventional cytogenetic analysis were performed in 207 children with B-ALL.

RESULTSIn 207 B-ALL children, the three-probe FISH panel assay showed that 151 cases carried genetic aberrancies with a positive rate of 72.9%, including 44 cases with typical positive signal patterns and 148 cases with atypical signal patterns (among them 41 cases have multiprobe abberancy). The conventional cytogenetic analysis detected 53 cases chromosomal abnormality with a positive rate of 25.6%.

CONCLUSIONThe detection rate of genetic abnormalities in newly- diagnosed pediatric B-ALL could be significantly improved by using three-probe FISH panel upon conventional cytogenetic analysis.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Infant ; Karyotyping ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics

OBJECTIVES: To study the clinical and prognostic significance of the preferentially expressed antigen of melanoma (PRAME) gene in the absence of specific fusion gene expression in children with B-lineage acute lymphoblastic leukemia (B-ALL). METHODS: A total of 167 children newly diagnosed with B-ALL were enrolled, among whom 70 were positive for the PRAME gene and 97 were negative. None of the children were positive for MLL-r, BCR/ABL, E2A/PBX1, or ETV6/RUNX1. The PRAME positive and negative groups were analyzed in terms of clinical features, prognosis, and related prognostic factors. RESULTS: Compared with the PRAME negative group, the PRAME positive group had a significantly higher proportion of children with the liver extending >6 cm below the costal margin (P<0.05). There was a significant reduction in the PRAME copy number after induction chemotherapy (P<0.05). In the minimal residual disease (MRD) positive group after induction chemotherapy, the PRAME copy number was not correlated with the MRD level (P>0.05). In the MRD negative group, there was also no correlation between them (P>0.05). The PRAME positive group had a significantly higher 4-year event-free survival rate than the PRAME negative group (87.5%±4.6% vs 73.5%±4.6%, P<0.05), while there was no significant difference between the two groups in the 4-year overall survival rate (88.0%±4.4% vs 85.3%±3.8%, P>0.05). The Cox proportional-hazards regression model analysis showed that positive PRAME expression was a protective factor for event-free survival rate in children with B-ALL (P<0.05). CONCLUSIONS Although the PRAME gene cannot be monitored as MRD, overexpression of PRAME suggests a good prognosis in B-ALL.
Acute Disease ; Antigens, Neoplasm/therapeutic use* ; Child ; Humans ; Neoplasm, Residual/diagnosis* ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/genetics* ; Prognosis

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

The aim of this study was to establish a method for quantitative detection of immunoglobulin heavy chain (IgH) gene rearrangements and to explore its clinical application in monitoring minimal residual disease (MRD) of acute lymphoid leukemia (ALL). Clonal IgH gene rearrangements in 51 patients with ALL at diagnosis were identified by multiplex PCR assay. PCR products were sequenced and pairwise in IMGT data base. Allele-specific oligonucleotides primers were designed complementary to the junctional region. The conservative JH primers combined with TaqMan probes were used to monitor the level of MRD in ALL patients. The sensitivity and specificity were assessed as well. The results indicated that out of all the 51 ALL patients, IgH rearrangements were identified in 21 cases, and 15 patients out of them were quantified. The slope of the standard curves was -3.1 to -3.9 and the correlation coefficients of all standard curves were > 0.98. The sensitivity was between 10(-4) and 10(-5), only one patient's sensitivity was 10(-3). Most of the quantitative range was less than 10(-4). The background's nonspecific amplification was detectable at a low level and had a little influence on results. 7 patients whose MRD level below 10(-3) kept complete remission and another 8 patients whose MRD level above 10(-3) had a higher relapse rate. It is concluded that the analysis of IgH gene rearrangements with RQ-PCR is a highly sensitive, specific and reliable method for accurate evaluation of MRD in ALL. The data indicates a high correlation between the level of IgH rearrangements and the prognosis in ALL patients.
Alleles ; DNA Primers ; genetics ; Female ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Male ; Neoplasm, Residual ; diagnosis ; genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics

Philadelphia chromosome-like acute lymphoblastic leukemia (Ph-like ALL) is a subtype of B-lineage ALL (B-ALL) that displays a gene expression profile (GEP) similar to Philadelphia chromosome-positive ALL (PhALL). It has a diverse range of genetic alterations that activate cytokine receptor genes and kinase signaling pathways, frequently accompanied by abnormal transcription factors related to lymphatic development. Children with Ph-like ALL account for 15% of children with high-risk B-ALL. It has adverse clinical features and a poor prognosis. Tyrosine kinase inhibitors combined with chemotherapy can significantly improve the prognosis of children with PhALL, suggesting that targeted therapy based on the molecular cytogenetic abnormalities of Ph-like ALL has good research prospects. This paper expounds the genetic alterations, pathogenesis, clinical features, diagnostic measures, and potential therapeutic approaches of Ph-like childhood ALL based on recent research progress in Ph-like ALL.
Humans ; Janus Kinase 2 ; genetics ; PAX5 Transcription Factor ; genetics ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; drug therapy ; genetics ; Proto-Oncogene Proteins c-abl ; genetics

OBJECTIVEThe study was aimed to investigate the feasibility and clinical significance of quantitative detection of minimal residual disease (MRD) in childhood acute lymphoblastic leukemia (ALL) by real-time quantitative polymerase chain reaction (RQ-PCR).

METHODSClonal IgH gene rearrangements of samples at diagnosis were identified by standard PCR assay with consensus primers. Monoclonal IgH gene rearrangements were analyzed using DNAPLOT software. Upstream primers were designed with the Primer Express software and allele specific oligonucleotide developed complementary to the V-D or D-J junction. Samples at diagnosis were serially diluted to generate the patient specific standard curves. RQ-PCR method was used to quantify the MRD of the follow up samples collected at five time points during chemotherapy. To check the quantity and quality of DNA, the investigators used RQ-PCR analysis for the albumin gene.

RESULTSTotally 16 monoclonal IgH gene rearrangements were identified from 34 patients with B-ALL. The analysis of the 16 monoclonal rearrangements showed that the most frequently used V segment was from V3 family and J segment from J4 and J6. The RQ-PCR sensitivity of 10(-4) to 10(-5) was mostly reached. Non-specific amplification was seen in 6 patients. The number of inserted and deleted nucleotides did not appear to be related to the sensitivity (P > 0.05). The correlation coefficients of all 16 standard curves were excellent (> or = 0.99). The mean slope of the standard curves was -3.4 +/- 0.37 and the mean intercept was 24.3 +/- 2.95. MRD analysis of follow up samples from the 16 patients showed an association between high degree of MRD and relapse. There was no apparent relationship between MRD degree at the end of induction chemotherapy and other high risk factors of ALL (P > 0.05).

CONCLUSIONThe study showed that the above approach with RQ-PCR was applicable to clinical detection of MRD in childhood ALL. Quantitative and dynamic study of MRD was of prognostic importance.

Child ; Gene Rearrangement, B-Lymphocyte ; Humans ; Neoplasm, Residual ; diagnosis ; genetics ; Polymerase Chain Reaction ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; drug therapy ; genetics ; Prognosis