The chromosome 4q21 gene (AF-4/FEL) is widely expressed in normal tissues and shows breakpoint diversity in t(4;11)(q21;q23) acute leukemia

The chromosomal translocation, t(4;11)(q21;q23), is the most common type of 11q23 chromosomal abnormality, being highly prevalent in infant acute leukemias and associated with a poor prognosis. The t(4;11) results in the fusion of an 11q23 gene (MLL, HRX, Htrx-1, or ALL-1) and a 4q21 gene (AF-4 or FEL). To further evaluate the 4q21 gene and its role in t(4;11) acute leukemia, we have cloned a 38-kb genomic region and mapped exons of the AF-4 gene. The 4q21 breakpoints in 19 cases of t(4;11) acute leukemia were analyzed by Southern analysis and pulsed- field gels. Seventeen of the 19 cases had breakpoints on chromosome 4q21 that were scattered in this 38 kb region. Expression of the AF-4 gene was studied in a total of 28 various nonhematopoietic, hematopoietic, and t(4;11) leukemic cell lines. The AF-4 gene was expressed in all cell lines as a major and a minor transcript. In addition to the normal transcripts, two fusion transcripts from the derivative 11 and derivative 4 chromosomes were identified in all t(4;11) cell lines except B1, which had only the der(11) transcript. These findings suggest that the breakpoints on 4q21 cluster over a broader area than do the breakpoints in the 11q23 gene, and that der(11) encodes the fusion RNA found consistently in leukemia cells.

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The chromosome 4q21 gene (AF-4/FEL) is widely expressed in normal tissues and shows breakpoint diversity in t(4;11)(q21;q23) acute leukemia

Blood ◽

10.1182/blood.v82.4.1080.1080 ◽

1993 ◽

Vol 82 (4) ◽

pp. 1080-1085 ◽

Cited By ~ 5

Author(s):

CS Chen ◽

JM Hilden ◽

J Frestedt ◽

PH Domer ◽

R Moore ◽

...

Keyword(s):

Acute Leukemia ◽

Cell Lines ◽

Chromosomal Translocation ◽

Leukemic Cell ◽

Genomic Region ◽

Acute Leukemias ◽

Normal Tissues ◽

Leukemic Cell Lines ◽

A Minor ◽

Chromosome 4Q21

Abstract The chromosomal translocation, t(4;11)(q21;q23), is the most common type of 11q23 chromosomal abnormality, being highly prevalent in infant acute leukemias and associated with a poor prognosis. The t(4;11) results in the fusion of an 11q23 gene (MLL, HRX, Htrx-1, or ALL-1) and a 4q21 gene (AF-4 or FEL). To further evaluate the 4q21 gene and its role in t(4;11) acute leukemia, we have cloned a 38-kb genomic region and mapped exons of the AF-4 gene. The 4q21 breakpoints in 19 cases of t(4;11) acute leukemia were analyzed by Southern analysis and pulsed- field gels. Seventeen of the 19 cases had breakpoints on chromosome 4q21 that were scattered in this 38 kb region. Expression of the AF-4 gene was studied in a total of 28 various nonhematopoietic, hematopoietic, and t(4;11) leukemic cell lines. The AF-4 gene was expressed in all cell lines as a major and a minor transcript. In addition to the normal transcripts, two fusion transcripts from the derivative 11 and derivative 4 chromosomes were identified in all t(4;11) cell lines except B1, which had only the der(11) transcript. These findings suggest that the breakpoints on 4q21 cluster over a broader area than do the breakpoints in the 11q23 gene, and that der(11) encodes the fusion RNA found consistently in leukemia cells.

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DIGE-Based Proteomic Analysis Identifies NPM/B23 and Nucleolin C23 as Overexpressed Proteins In Relapsed /Refractory Acute Leukemia

Blood ◽

10.1182/blood.v116.21.1711.1711 ◽

2010 ◽

Vol 116 (21) ◽

pp. 1711-1711

Author(s):

Jian Da Hu ◽

MinHui Lin ◽

TingBo Liu ◽

Jing Li ◽

XinJi Chen ◽

...

Keyword(s):

Treatment Outcome ◽

Multidrug Resistance ◽

Acute Leukemia ◽

Western Blot ◽

Cell Lines ◽

Molecular Mechanisms ◽

Expression Profiles ◽

Leukemia Cell ◽

Leukemic Cell ◽

Leukemic Cell Lines

Abstract Abstract 1711 Resistance to chemotherapy is a challenge in treatment of acute leukemia. Although the classic multidrug resistance (MDR) phenotype is often characterized by expression of drug efflux pump P-glycoprotein or by multidrug resistance-associated proteins, precise molecular mechanisms are largely unknown. To investigate novel protein changes involved in resistance mechanism, protein expression profiles between human myeloid leukemia HL-60 cell lines and adriamycin- resistant HL-60 cell lines (HL-60/ADR) was compared, which was based on a differential proteomic approach — 2 dimensional difference in gel Electrophoresis(2D-DIGE) followed by mass spectrometry (MALDI-TOF-MS) and complemented by western blot validation. 16 protein spots were identified as being differentially expressed (> 1.2 fold change and p≤ 0.05) between above two cell lines, among which 13 protein spots were identified as up-regulated and 3 as down-regulated in the HL-60/ ADR cell line. Proteins found to have higher abundance levels in the resistant HL-60/ADR cells included enzymes, proteins and oncogenes related to signal transduction, protein synthesis, cell growth regulation and metabolism. 3 lower abundance proteins are related to transcription. From 16 proteins, 2 proteins, nucleophosmin B23 (NPM B23)and nucleolin C23, were selected and verified in leukemia cell lines and primary leukemia samples by western blot. Compared to healthy control samples, which showed no expressions of these 2 proteins, leukemic cell lines revealed an obvious up-regulation of B23 and C23. Moreover, significantly higher expressions of B23 and C23 were found in 3 resistant leukemic cell lines, HL-60/ADR, K562/ADR and KG01 cells, compared to the parent HL-60 and K562 cells, and other leukemic cell lines. In de novo leukemia samples, 43.8%(35/80) expressed B23 and C23 proteins, 37.9% (22/58) AML and 59.1% (13/22) ALL respectively. Meanwhile, concomitant expression of B23 and C23, both positive or negative, was noted in 97%(79/80)patients. Over-expressions of B23 and C23 were observed in 68.8% relapased/refractory leukemia patients. With regard to treatment outcome,among those patients who achieved ongoing CR, fewer patients expressed 2 proteins, only 13.35% (7/52) AML and 46%(7/15) ALL respectively. It implicated that B23 and C23 may be involved in drug resistance and be useful in assessing treatment outcome and prognosis of leukemia. To a conclusion, these results provide a novel clue for the molecular mechanism of MDR and suggest that B23 and C23 are prognostic indicators for leukemia. Disclosures: No relevant conflicts of interest to declare.

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Abnormalities of the retinoblastoma gene in the pathogenesis of acute leukemia

Blood ◽

10.1182/blood.v78.12.3259.3259 ◽

1991 ◽

Vol 78 (12) ◽

pp. 3259-3268 ◽

Cited By ~ 85

Author(s):

HG Ahuja ◽

PS Jat ◽

A Foti ◽

M Bar-Eli ◽

MJ Cline

Keyword(s):

Acute Leukemia ◽

Protein Expression ◽

Blast Crisis ◽

Philadelphia Chromosome ◽

Leukemic Cell ◽

Monocytic Differentiation ◽

Lymphoid Leukemia ◽

Acute Leukemias ◽

Leukemic Cell Lines ◽

Rb Gene

Abstract The retinoblastoma-susceptibility (Rb) gene is an antioncogene that is frequently altered in retinoblastomas, sarcomas, and some epithelial tumors. We examined the structure of the Rb gene by Southern blotting in 215 cases of leukemias and lymphomas of diverse phenotype and in 15 leukemic cell lines. In selected cases Rb protein expression was examined with specific monoclonal antibodies. Structural abnormalities of the Rb gene with absent protein expression were frequent in all types of human acute leukemia, but were particularly common (27% incidence) in M4 and M5 myeloid leukemia with monocytic differentiation and in Philadelphia chromosome (Ph1)-positive leukemia of lymphoid phenotype (11% to 29% incidence). Changes in Rb were observed early in the transition to acute leukemia in cases of myelodysplastic syndrome and in the accelerated phase of chronic myelocytic leukemia in transition to blast crisis. In one case, molecular changes in Rb could be correlated with leukemia remission and relapse. We conclude that the Rb antioncogene is commonly involved in the evolution of human acute leukemias, particularly in those of a monocytic phenotype and in lymphoid leukemia in which there is an antecedent alteration of the Ph1 chromosome.

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Growth factor requirements of childhood acute leukemia: establishment of GM-CSF-dependent cell lines

Blood ◽

10.1182/blood.v70.1.192.bloodjournal701192 ◽

1987 ◽

Vol 70 (1) ◽

pp. 192-199 ◽

Cited By ~ 1

Author(s):

B Lange ◽

M Valtieri ◽

D Santoli ◽

D Caracciolo ◽

F Mavilio ◽

...

Keyword(s):

Growth Factor ◽

Acute Leukemia ◽

Cell Lines ◽

Leukemic Cell ◽

Media Analysis ◽

Leukemic Cells ◽

Gm Csf ◽

Leukemic Cell Lines ◽

Dependent Cell ◽

Childhood Acute Leukemia

Eight permanent cell lines were established from cells of 50 consecutive patients with childhood acute leukemia. Three cell lines required growth factor-containing conditioned media. Analysis using blocking antisera and recombinant granulocytic macrophage (GM) colony- stimulating factor (CSF) identified GM-CSF as a growth factor required to establish the latter three cell lines and necessary for their continuous proliferation in chemically defined medium. Two of the GM- CSF-dependent cell lines were derived from patients with undifferentiated T- and a biphenotypic B-myelomonocytic leukemia, which suggests that GM-CSF might maintain proliferation of leukemias originating from immature progenitor cells. Cytogenetic analysis indicated that all established leukemic cell lines were aneuploid, with six lines containing chromosomal alterations related to those observed in the leukemic cells of the patient. Two patients did not have an abnormal clone identified in the marrow but did yield an aneuploid cell line. These studies indicate that GM-CSF-dependent leukemic cell lines can be established in a fraction of childhood leukemia. These cell lines lend themselves to studies aimed at the evaluation in vitro of the role of growth factors in controlling proliferation and differentiation of leukemic cells.

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Abnormalities of the retinoblastoma gene in the pathogenesis of acute leukemia

Blood ◽

10.1182/blood.v78.12.3259.bloodjournal78123259 ◽

1991 ◽

Vol 78 (12) ◽

pp. 3259-3268 ◽

Cited By ~ 2

Author(s):

HG Ahuja ◽

PS Jat ◽

A Foti ◽

M Bar-Eli ◽

MJ Cline

Keyword(s):

Acute Leukemia ◽

Protein Expression ◽

Blast Crisis ◽

Philadelphia Chromosome ◽

Leukemic Cell ◽

Monocytic Differentiation ◽

Lymphoid Leukemia ◽

Acute Leukemias ◽

Leukemic Cell Lines ◽

Rb Gene

The retinoblastoma-susceptibility (Rb) gene is an antioncogene that is frequently altered in retinoblastomas, sarcomas, and some epithelial tumors. We examined the structure of the Rb gene by Southern blotting in 215 cases of leukemias and lymphomas of diverse phenotype and in 15 leukemic cell lines. In selected cases Rb protein expression was examined with specific monoclonal antibodies. Structural abnormalities of the Rb gene with absent protein expression were frequent in all types of human acute leukemia, but were particularly common (27% incidence) in M4 and M5 myeloid leukemia with monocytic differentiation and in Philadelphia chromosome (Ph1)-positive leukemia of lymphoid phenotype (11% to 29% incidence). Changes in Rb were observed early in the transition to acute leukemia in cases of myelodysplastic syndrome and in the accelerated phase of chronic myelocytic leukemia in transition to blast crisis. In one case, molecular changes in Rb could be correlated with leukemia remission and relapse. We conclude that the Rb antioncogene is commonly involved in the evolution of human acute leukemias, particularly in those of a monocytic phenotype and in lymphoid leukemia in which there is an antecedent alteration of the Ph1 chromosome.

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Carbohydrate antigen profiles of human erythroleukemia cell lines HEL and K562

Blood ◽

10.1182/blood.v62.6.1230.1230 ◽

1983 ◽

Vol 62 (6) ◽

pp. 1230-1241 ◽

Cited By ~ 23

Author(s):

R Kannagi ◽

T Papayannopoulou ◽

B Nakamoto ◽

NA Cochran ◽

T Yokochi ◽

...

Keyword(s):

Cell Lines ◽

Leukemic Cell ◽

K562 Cells ◽

Carbohydrate Antigen ◽

Carbohydrate Antigens ◽

Leukemic Cell Lines ◽

Hel Cells ◽

Erythroleukemia Cell ◽

A Minor ◽

I Cells

Abstract The expression of major carbohydrate antigens carried by polylactosaminyl chains in human erythroleukemia cell lines, K562 and HEL, was investigated by applying monoclonal antibodies recognizing specific carbohydrate determinants. The two cell lines showed common differences in their glycolipid compositions: (1) the presence of significant amounts of ganglio-series glycolipids, which are absent in normal erythrocytes; and (2) a remarkable reduction in the amount of globo-series glycolipids, which are the major glycolipids in normal human erythrocytes. A variety of differences were also detected in the carbohydrate antigens carried by lacto-series glycolipids and glycoproteins having related carbohydrate chains. K562 cells were i+H- X+, with a minor population of I+ cells. HEL cells were I-i+H+X-. The presence of the I+ population in K562 cells is particularly noteworthy, since I-antigen is characteristic of adult mature erythrocytes and is absent in most human leukemic cell lines. Several clones showing I+, I+/-i+/-, or I-i+ specificities were isolated from K562 cells by cloning in either methylcellulose media or limiting dilution, and I+ and I- cells were sorted by FACS fluorometer. HEL cells and these K562 clones provide a useful experimental model for studying the biologic significance and enzymatic control in expression of cell surface polylactosamines.

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Growth factor requirements of childhood acute leukemia: establishment of GM-CSF-dependent cell lines

Blood ◽

10.1182/blood.v70.1.192.192 ◽

1987 ◽

Vol 70 (1) ◽

pp. 192-199 ◽

Cited By ~ 142

Author(s):

B Lange ◽

M Valtieri ◽

D Santoli ◽

D Caracciolo ◽

F Mavilio ◽

...

Keyword(s):

Growth Factor ◽

Acute Leukemia ◽

Cell Lines ◽

Leukemic Cell ◽

Media Analysis ◽

Leukemic Cells ◽

Gm Csf ◽

Leukemic Cell Lines ◽

Dependent Cell ◽

Childhood Acute Leukemia

Abstract Eight permanent cell lines were established from cells of 50 consecutive patients with childhood acute leukemia. Three cell lines required growth factor-containing conditioned media. Analysis using blocking antisera and recombinant granulocytic macrophage (GM) colony- stimulating factor (CSF) identified GM-CSF as a growth factor required to establish the latter three cell lines and necessary for their continuous proliferation in chemically defined medium. Two of the GM- CSF-dependent cell lines were derived from patients with undifferentiated T- and a biphenotypic B-myelomonocytic leukemia, which suggests that GM-CSF might maintain proliferation of leukemias originating from immature progenitor cells. Cytogenetic analysis indicated that all established leukemic cell lines were aneuploid, with six lines containing chromosomal alterations related to those observed in the leukemic cells of the patient. Two patients did not have an abnormal clone identified in the marrow but did yield an aneuploid cell line. These studies indicate that GM-CSF-dependent leukemic cell lines can be established in a fraction of childhood leukemia. These cell lines lend themselves to studies aimed at the evaluation in vitro of the role of growth factors in controlling proliferation and differentiation of leukemic cells.

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Carbohydrate antigen profiles of human erythroleukemia cell lines HEL and K562

Blood ◽

10.1182/blood.v62.6.1230.bloodjournal6261230 ◽

1983 ◽

Vol 62 (6) ◽

pp. 1230-1241 ◽

Cited By ~ 1

Author(s):

R Kannagi ◽

T Papayannopoulou ◽

B Nakamoto ◽

NA Cochran ◽

T Yokochi ◽

...

Keyword(s):

Cell Lines ◽

Leukemic Cell ◽

K562 Cells ◽

Carbohydrate Antigen ◽

Carbohydrate Antigens ◽

Leukemic Cell Lines ◽

Hel Cells ◽

Erythroleukemia Cell ◽

A Minor ◽

I Cells

The expression of major carbohydrate antigens carried by polylactosaminyl chains in human erythroleukemia cell lines, K562 and HEL, was investigated by applying monoclonal antibodies recognizing specific carbohydrate determinants. The two cell lines showed common differences in their glycolipid compositions: (1) the presence of significant amounts of ganglio-series glycolipids, which are absent in normal erythrocytes; and (2) a remarkable reduction in the amount of globo-series glycolipids, which are the major glycolipids in normal human erythrocytes. A variety of differences were also detected in the carbohydrate antigens carried by lacto-series glycolipids and glycoproteins having related carbohydrate chains. K562 cells were i+H- X+, with a minor population of I+ cells. HEL cells were I-i+H+X-. The presence of the I+ population in K562 cells is particularly noteworthy, since I-antigen is characteristic of adult mature erythrocytes and is absent in most human leukemic cell lines. Several clones showing I+, I+/-i+/-, or I-i+ specificities were isolated from K562 cells by cloning in either methylcellulose media or limiting dilution, and I+ and I- cells were sorted by FACS fluorometer. HEL cells and these K562 clones provide a useful experimental model for studying the biologic significance and enzymatic control in expression of cell surface polylactosamines.

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Repositioning of Difluorinated Propanediones as Inhibitors of Histone Methyltransferases and their Biological Evaluation in Human Leukemic Cell Lines

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520618666180404125721 ◽

2019 ◽

Vol 18 (13) ◽

pp. 1892-1899 ◽

Cited By ~ 1

Author(s):

Tanushree Pal ◽

Asmita Sharda ◽

Bharat Khade ◽

C. Sinha Ramaa ◽

Sanjay Gupta

Keyword(s):

Cell Lines ◽

Physiological Role ◽

Leukemic Cell ◽

Docking Studies ◽

Biological Evaluation ◽

Leukemic Cell Lines ◽

Histone Lysine ◽

Histone Lysine Methyltransferase ◽

Subsequent Decrease

Background: At present, ‘pharmaco-epigenomics’ constitutes the hope in cancer treatment owing to epigenetic deregulation- a reversible process and playing a role in malignancy. Objective: Chemotherapy has many limitations like host-tissue toxicity, drug resistance. Hence, it is imperative to unearth targets to better treat cancer. Here, we intend to repurpose a set of our previously synthesized difluorinated Propanediones (PR) as Histone lysine Methyltransferase inhibitors (HMTi). Methods: The cell lines of leukemic origin viz. histiocytic lymphoma (U937) and acute T-cell leukemia (JURKAT) were treated with PR-1 to 7 after docking studies with active pocket of HMT. The cell cycle analysis, in vitro methylation and cell proliferation assays were carried out to delineate their physiological role. Results: A small molecule PR-4, at 1 and 10µM, has shown to alter the methylation of histone H3 and H4 in both cell lines. Also, treatment shows an increase in G2/M population and a subsequent decrease in the G0/G1 population in U937. In JURKAT, an increase in both G2/M and S phase population was observed. The sub-G1 population showed a steady rise with increase in dose and prolonged time intervals in U937 and JURKAT cell lines. In SRB assay, the PR showed a cell growth of 42.6 and 53.4% comparable to adriamycin; 44.5 and 53.2% in U937 and JURKAT, respectively. The study suggests that PR-4 could emerge as a potential HMT inhibitor. Conclusion: The molecule PR-4 could be a lead in developing more histone lysine methyltransferases inhibitors with potential to be pro-apoptotic agents.

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