Abstract:
:Single-colony karyotyping (SCK) and reverse-transcription polymerase chain reaction (RT-PCR) are two increasingly used techniques for the quantification of leukemic colonies generated by chronic myelogenous leukemia (CML) cell fractions purged or selected in vitro. Recently, the existence of Philadelphia (Ph) chromosome positive progenitors with a silent BCR-ABL gene has been reported, thus raising concerns on the use of RT-PCR for detecting BCR-ABL positive progenitors. In order to investigate this issue further, colonies (n = 204) generated by mononuclear (MNC) or CD34+ CML cells were individually harvested, divided into two aliquots and analyzed both at the cytogenetic level to detect the Ph chromosome, and the molecular level to detect BCR-ABL transcripts. The mean (+/- s.d.) percentages of colonies analyzable by either SCK or RT-PCR were 74 +/- 16% and 86 +/- 16%, respectively. A significant percentage of colonies (67 +/- 19%) could be successfully analyzed by both SCK and RT-PCR. Although the majority of these colonies (97 +/- 5%) were Ph-positive and BCR-ABL-positive, a negligible percentage (4%) of progenitors were Ph-positive but BCR-ABL-negative. In order to test the influence of colony size on the outcome of molecular analysis, the efficiency of our RT-PCR assay in detecting BCR-ABL transcripts was investigated by means of experiments in which the number of cells used to start RNA extraction was serially reduced. These experiments showed that at least 150 cells were necessary to achieve a reproducible amplification of BCR-ABL transcripts. By correlating the size of harvested colonies with the outcome of molecular analysis, it was evident that BCR-ABL-negative but Ph-positive colonies represented false negative results occurring when a number of leukemic cells below the detection limit of our RT-PCR assay was analyzed. In conclusion, our data demonstrate that individual CML colonies grown in semisolid culture assays can be indifferently analyzed by SCK or RT-PCR, and support an extensive use of a carefully standardized RT-PCR assay to estimate the leukemic burden within samples which have been purged and selected in vitro.
journal_name
Leukemiajournal_title
Leukemiaauthors
Savoldo B,Sammarelli G,Dotti G,Garau D,Regazzi E,Cilloni D,Tabilio A,Rizzoli V,Carlo-Stella Cdoi
10.1038/sj.leu.2400942subject
Has Abstractpub_date
1998-03-01 00:00:00pages
434-40issue
3eissn
0887-6924issn
1476-5551journal_volume
12pub_type
杂志文章相关文献
LEUKEMIA文献大全abstract::Human promonocytic cell line U937 cells can be induced to differentiate into macrophages by treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA). TPA treatment induced the expression of the monocytic differentiation markers CD11b and CD36, with concomitant morphological changes. Moreover, TPA enhanced reactive ox...
journal_title:Leukemia
pub_type: 杂志文章
doi:10.1038/leu.2008.353
更新日期:2009-04-01 00:00:00
abstract::The FMS-like tyrosine kinase-3 (FLT3) gene is the most commonly mutated gene in acute myeloid leukemia (AML), and patients carrying internal tandem duplication (ITD) mutations have a poor prognosis. Long-term inhibition of FLT3 activity in these patients has been elusive. To provide a more complete understanding of FL...
journal_title:Leukemia
pub_type: 杂志文章
doi:10.1038/leu.2016.284
更新日期:2017-03-01 00:00:00
abstract::Clinical characteristics, treatment response and outcome were evaluated in children with Down's syndrome (DS) and acute lymphoblastic leukemia (ALL) as compared to other children with ALL (NDS). Sixty-one DS and 4049 NDS patients, receiving intensive antileukemic treatment during four consecutive trials (ALL-BFM 81, 8...
journal_title:Leukemia
pub_type: 杂志文章
doi:10.1038/sj.leu.2400989
更新日期:1998-05-01 00:00:00
abstract::Cytogenetic and fluorescence in situ hybridization studies have shown the presence of telomeric repeats in translocation present in three patients with hematopoietic malignancies. One had jumping translocations, involving 1q12 and 2q, 16p, and 19q. These sequences were detected by FISH only in derivative chromosomes t...
journal_title:Leukemia
pub_type: 杂志文章
doi:10.1038/sj.leu.2401876
更新日期:2000-09-01 00:00:00
abstract::Immunophenotypes and Ig gene rearrangements were investigated in 12 patients with a variant form of hairy cell leukemia (HCL) termed HCL-Japanese variant (HCL-J), and in an HCL-J-derived cell line. The leukemic cells of HCL-J characteristically showed the phenotype of CD20+, CD5-, CD10-, CD11c+, CD22+, CD24- and CD25-...
journal_title:Leukemia
pub_type: 杂志文章
doi:
更新日期:1996-08-01 00:00:00
abstract::The translocation t(11;20)(p15;q11) was found as the sole acquired clonal chromosome abnormality in two patients with acute myeloid leukemia. The bone marrow morphology in both cases corresponded to the M2 subtype of the French-American-British (FAB) classification. None of the patients achieved complete remission, an...
journal_title:Leukemia
pub_type: 杂志文章
doi:
更新日期:1988-07-01 00:00:00
abstract::Reciprocal rearrangements of the MLL gene are among the most common chromosomal abnormalities in both Acute Lymphoblastic and Myeloid Leukemia. The MLL gene, located on the 11q23 chromosomal band, is involved in more than 40 recurrent translocations. In the present study, we describe the development and validation of ...
journal_title:Leukemia
pub_type: 杂志文章
doi:10.1038/sj.leu.2403439
更新日期:2004-09-01 00:00:00
abstract::T-cell acute lymphoblastic leukemia (T-ALL) is associated with chromosomal aberrations characterized by juxtaposition of proto-oncogenes to T-cell receptor gene loci (TCR), resulting in the deregulated transcription of these proto-oncogenes. Here, we describe the molecular characterization of a novel chromosomal aberr...
journal_title:Leukemia
pub_type: 杂志文章
doi:10.1038/sj.leu.2403619
更新日期:2005-02-01 00:00:00
abstract::The role of transforming growth factor-β (TGFβ) signaling in embryological development and tissue homeostasis has been thoroughly characterized. Its canonical downstream cascade is well known, even though its true complexity and other non-canonical pathways are still being explored. TGFβ signaling has been described a...
journal_title:Leukemia
pub_type: 杂志文章,评审
doi:10.1038/s41375-019-0420-1
更新日期:2019-05-01 00:00:00
abstract::Neomorphic mutations in isocitrate dehydrogenase 1 (IDH1) are frequently found in several human cancer types including acute myeloid leukemia (AML) and lead to the production of high levels of the oncometabolite (R)-2-hydroxyglutarate (R-2HG). Here we report the characterization of BAY1436032, a novel pan-mutant IDH1 ...
journal_title:Leukemia
pub_type: 杂志文章
doi:10.1038/leu.2017.46
更新日期:2017-10-01 00:00:00
abstract::Chronic lymphocytic leukemia (CLL) cells with aggressive clinical properties express lipoprotein lipase (LPL), which generates activating ligands for the nuclear receptor peroxisome proliferator activated receptor (PPAR)α and allows fatty acids to be used as fuel. However, the role of PPARα in CLL is unclear. PPARα wa...
journal_title:Leukemia
pub_type: 杂志文章
doi:10.1038/leu.2012.329
更新日期:2013-04-01 00:00:00
abstract::In order to investigate the frequency of HHV-8 in MM patients from another geographic location, we obtained fresh bone marrow (BM) biopsies from Turkish patients with MM (n = 21), monoclonal gammopathy of undetermined significance (MGUS) (n = 2), plasmacytoma (n = 1) with BM plasma cell infiltration, various hematolog...
journal_title:Leukemia
pub_type: 杂志文章
doi:10.1038/sj.leu.2402190
更新日期:2001-08-01 00:00:00
abstract::Despite the presence of tumor antigens, the paucity of clinically significant T-cell mediated immune responses against human tumors is striking. This may, in part, be because of the inability of cancer cells to function as efficient antigen-presenting cells. For full activation, T cells must receive two signals delive...
journal_title:Leukemia
pub_type: 杂志文章,评审
doi:
更新日期:1997-05-01 00:00:00
abstract::Granulocyte-macrophage colony-stimulating factor (GM-CSF) plays a critical role in growth and differentiation of myeloid cells. We previously reconstituted high affinity human GM-CSF receptor (hGM-CSFR) in a proB cell line BA/F3 by cotransfecting alpha and beta chain cDNA clones and showed that the reconstituted recep...
journal_title:Leukemia
pub_type: 杂志文章
doi:
更新日期:1993-08-01 00:00:00
abstract::The impact of ten-eleven-translocation 2 (TET2) mutations on response to azacitidine (AZA) in MDS has not been reported. We sequenced the TET2 gene in 86 MDS and acute myeloid leukemia (AML) with 20-30% blasts treated by AZA, that is disease categories wherein this drug is approved by Food and Drug Administration (FDA...
journal_title:Leukemia
pub_type: 杂志文章
doi:10.1038/leu.2011.71
更新日期:2011-07-01 00:00:00
abstract::Blockade of tumor necrosis factor (TNF)alpha by a soluble TNF receptor fusion protein (etanercept; Enbrel) improved in vitro hemopoiesis from the marrow of patients with myelodysplastic syndrome (MDS). Therefore, we enrolled 14 MDS patients (4 RA, 2 RARS, 6 RAEB, 2 CMML), 44-80 (median 60) years old, in a pilot trial....
journal_title:Leukemia
pub_type: 临床试验,杂志文章
doi:10.1038/sj.leu.2402356
更新日期:2002-02-01 00:00:00
abstract::Translocation (15;17)(q22;q12-q21) is a chromosome aberration specifically found in acute promyelocytic leukemia (APL), that generates a chimeric gene between the promyelocytic leukemia (PML) gene on chromosome 15 and the retinoic acid receptor alpha (RARA) gene, on chromosome 17. In the course of molecular investigat...
journal_title:Leukemia
pub_type: 杂志文章
doi:
更新日期:1993-01-01 00:00:00
abstract::Multiple myeloma (MM) is a plasma cell tumor marked by clonal evolution and preceded by a premalignant stage, which progresses via molecular pathway deregulation, including MYC activation. This activation relates to translocation or gain of the MYC locus and deregulation of upstream pathways such as IRF4, DIS3/LIN28B/...
journal_title:Leukemia
pub_type: 杂志文章,评审
doi:10.1038/s41375-018-0036-x
更新日期:2018-06-01 00:00:00
abstract::Mutations of signal transducing molecules such as Ras have been shown to confer a growth advantage in leukaemic blasts and contribute to the pathogenesis of the disease. Alterations of signal transducing molecules other than Ras may play a role in leukaemogenesis. Knowledge of such mutations could also further our und...
journal_title:Leukemia
pub_type: 杂志文章
doi:
更新日期:1994-09-01 00:00:00
abstract::Deletions from the derivative chromosome 9, der(9), of the translocation, t(9;22)(q34;q11), at the site of the ABL/BCR fusion gene, have been demonstrated by fluorescence in situ hybridisation (FISH), in both Philadelphia chromosome (Ph)-positive chronic myeloid leukaemia (CML) and acute lymphoblastic leukaemia (ALL)....
journal_title:Leukemia
pub_type: 杂志文章
doi:10.1038/sj.leu.2403629
更新日期:2005-04-01 00:00:00
abstract::Engagement of NKG2D by their ligands (NKG2D-L), as the human major histocompatibility complex class I-related molecules MIC-A and the UL16-binding proteins, on cytolytic lymphocytes leads to the enhancement of antitumour effector functions. These ligands are missing or expressed at very low levels on leukaemic cells; ...
journal_title:Leukemia
pub_type: 杂志文章
doi:10.1038/leu.2008.354
更新日期:2009-04-01 00:00:00
abstract::The chromosomal inversion (16)(p13q22), which is associated with the M4-eosinophilia subtype of human acute myeloid leukemia, causes the fusion of two distinct genes. The polypeptide encoded by the chimeric gene, PEBP2p/CBFp-SMMHC, retains the ability to interact with, and dominantly interfere with the function of pro...
journal_title:Leukemia
pub_type: 杂志文章
doi:10.1038/sj.leu.2401821
更新日期:2000-07-01 00:00:00
abstract::Bruton's tyrosine kinase (BTK) inhibitor is a promising novel agent that has potential efficiency in B-cell malignancies. It took approximately 20 years from target discovery to new drug approval. The first-in-class drug ibrutinib creates possibilities for an era of chemotherapy-free management of B-cell malignancies,...
journal_title:Leukemia
pub_type: 杂志文章,评审
doi:10.1038/s41375-020-01072-6
更新日期:2020-10-29 00:00:00
abstract::We describe two new human leukemia cell lines, MOLM-13 and MOLM-14, established from the peripheral blood of a patient at relapse of acute monocytic leukemia, FAB M5a, which had evolved from myelodysplastic syndrome (MDS). Both cell lines express monocyte-specific esterase (MSE) and MLL-AF9 fusion mRNA. Gene fusion is...
journal_title:Leukemia
pub_type: 杂志文章
doi:10.1038/sj.leu.2400768
更新日期:1997-09-01 00:00:00
abstract::In vitro enzymatic amplification and direct sequencing were used to detect and characterize t(14;18) recombination junctions in peripheral blood and bone marrow mononuclear cell preparations from patients with follicular lymphoma in remission. Samples from 24/44 patients were found to be positive for translocations in...
journal_title:Leukemia
pub_type: 杂志文章
doi:
更新日期:1991-07-01 00:00:00
abstract::Basic fibroblast growth factor (bFGF) is a pleiotropic cytokine which has recently been shown to delay fludarabine-induced apoptosis in B cell chronic lymphocytic leukemia (B-CLL) cells. To investigate the potential mechanism of bFGF-mediated delay of apoptosis, two EBV-transformed B prolymphocytic cell lines (JVM-2, ...
journal_title:Leukemia
pub_type: 杂志文章
doi:10.1038/sj.leu.2400556
更新日期:1997-02-01 00:00:00
abstract::As typical disorders of the elderly, myelodysplastic syndromes (MDSs) are relatively unusual in childhood. Nevertheless, up to 17% of cases of pediatric acute myeloid leukemia may have a preleukemic phase. In young patients, the goal of treatment is eradication of the preleukemic malignant clone and reconstitution of ...
journal_title:Leukemia
pub_type: 杂志文章
doi:
更新日期:1994-05-01 00:00:00
abstract::We investigated the hypothesis that gemtuzumab ozogamicin (GO), an anti-CD33 immunotoxin would improve the efficacy of fludarabine/melphalan as a preparative regimen for allogeneic hematopoietic stem cell transplantation (HSCT) in a phase I/II trial. Toxicity was defined as grades III-IV organ damage, engraftment fail...
journal_title:Leukemia
pub_type: 杂志文章
doi:10.1038/sj.leu.2405014
更新日期:2008-02-01 00:00:00
abstract::To clarify the clinical and biological significance of beta 2-microglobulin (beta 2-M) in serum of adult T cell leukemia (ATL) associated with human lymphotropic virus type-I (HTLV-I), beta 2-M was measured in 52 patients with ATL (acute ATL, 35 patients; lymphoma ATL, two patients; chronic ATL, 12 patients; smolderin...
journal_title:Leukemia
pub_type: 杂志文章
doi:
更新日期:1995-04-01 00:00:00
abstract::Bone mineral density (BMD) and biochemical markers of bone metabolism were analyzed in 67 adults with ALL (n = 27), AML (n = 14), MDS (n = 6) and CML (n = 20) before and after allogeneic stem cell transplantation (SCT). Median age was 36 years (17-56). Twenty-six out of 53 patients (49%) had osteopenia and osteoporosi...
journal_title:Leukemia
pub_type: 临床试验,杂志文章
doi:10.1038/sj.leu.2402264
更新日期:2001-11-01 00:00:00