scholarly journals Erythropoietin-induced stimulation of differentiation and proliferation in J2E cells is not mimicked by chemical induction

Blood ◽  
1992 ◽  
Vol 80 (2) ◽  
pp. 412-419 ◽  
Author(s):  
SJ Busfield ◽  
SP Klinken
Keyword(s):  
Cell Line ◽  
Messenger Rna ◽  
Cellular Proliferation ◽  
Erythroid Cell ◽  
Chemical Induction ◽  
Chemically Induced ◽  
Proliferation And Differentiation ◽  
Differentiation And Proliferation ◽  
Sudden Decrease ◽  
Physiologic Stimulus

Abstract The J2E cell line is a novel erythroid cell line that differentiates in response to erythropoietin (Epo), the physiologic stimulus for erythropoiesis. After exposure to Epo, the cells synthesize hemoglobin, and we show here that this process is tightly linked to increases in cellular proliferation and DNA synthesis. The hormone-induced terminal differentiation also results in morphologic alterations and the accumulation of transcripts for alpha, beta maj, and beta min globins. c-myc messenger RNA levels increase rapidly after exposure to Epo and precede the increase in cell division, while c-myb undergoes a transient decrease. Differentiation of J2E cells can also be achieved with sodium butyrate, but, in contrast with Epo, this is associated with a retardation of replication and a sudden decrease in c-myc levels. These results show that, in this system, chemically induced differentiation differs from terminal maturation promoted by Epo and that the processes of proliferation and differentiation in J2E cells can be uncoupled.

scholarly journals Erythropoietin-induced stimulation of differentiation and proliferation in J2E cells is not mimicked by chemical induction

Blood ◽  
1992 ◽  
Vol 80 (2) ◽  
pp. 412-419 ◽  
Author(s):  
SJ Busfield ◽  
SP Klinken
Keyword(s):  
Cell Line ◽  
Messenger Rna ◽  
Cellular Proliferation ◽  
Erythroid Cell ◽  
Chemical Induction ◽  
Chemically Induced ◽  
Proliferation And Differentiation ◽  
Differentiation And Proliferation ◽  
Sudden Decrease ◽  
Physiologic Stimulus

The J2E cell line is a novel erythroid cell line that differentiates in response to erythropoietin (Epo), the physiologic stimulus for erythropoiesis. After exposure to Epo, the cells synthesize hemoglobin, and we show here that this process is tightly linked to increases in cellular proliferation and DNA synthesis. The hormone-induced terminal differentiation also results in morphologic alterations and the accumulation of transcripts for alpha, beta maj, and beta min globins. c-myc messenger RNA levels increase rapidly after exposure to Epo and precede the increase in cell division, while c-myb undergoes a transient decrease. Differentiation of J2E cells can also be achieved with sodium butyrate, but, in contrast with Epo, this is associated with a retardation of replication and a sudden decrease in c-myc levels. These results show that, in this system, chemically induced differentiation differs from terminal maturation promoted by Epo and that the processes of proliferation and differentiation in J2E cells can be uncoupled.

Hmgb3 Induces the Differentiation of Uterine Stromal Cells Through Targeting Ptn

2018 ◽  
Vol 26 (7) ◽  
pp. 891-899
Author(s):  
Kai Wang ◽  
Yun-Hou Yin ◽  
Zhan-Qing Yang ◽  
Hai-Fan Yu ◽  
Yu-Si Wang ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Messenger Rna ◽  
Cellular Proliferation ◽  
Physiological Role ◽  
High Mobility ◽  
Ovariectomized Mice ◽  
Dynamic Expression ◽  
Proliferation And Differentiation ◽  
Reliable Marker

Uterine decidualization is crucial for placenta formation and pregnancy maintenance. Although previous studies have reported that high mobility group box 3 (Hmgb3) is involved in the regulation of cellular proliferation and differentiation, little is known regarding its physiological role in uterine decidualization. Here, in situ hybridization result exhibited a dynamic expression pattern of Hmgb3 messenger RNA (mRNA) during early gestation, and it was mainly localized to the decidua on days 6 to 8 of gestation. Consistently, elevated Hmgb3 expression was noted in the decidualizing stromal cells after intraluminal oil infusion. In uterine luminal epithelium of ovariectomized mice, estrogen induced the accumulation of Hmgb3 mRNA, which was dependent on the existence of implanting blastocyst. Simultaneously, Hmgb3 could stimulate the proliferation of uterine stromal cells and promote the expression of Prl8a2, a reliable marker for stromal cell differentiation. Further analysis evidenced that Hmgb3 might modulate the expression of pleiotropin (Ptn) in uterine stromal cells. Moreover, silencing of Ptn could impede the upregulation of Prl8a2 elicited by Hmgb3 overexpression, while overexpression of Ptn reversed the repressive effects of Hmgb3 siRNA on Prl8a2 expression. Collectively, Hmgb3 may direct uterine decidualization through targeting Ptn.

scholarly journals KU 812: a pluripotent human cell line with spontaneous erythroid terminal maturation

Blood ◽  
1989 ◽  
Vol 73 (7) ◽  
pp. 2003-2013
Author(s):  
M Nakazawa ◽  
MT Mitjavila ◽  
N Debili ◽  
N Casadevall ◽  
P Mayeux ◽  
...  
Keyword(s):  
Cell Line ◽  
Messenger Rna ◽  
Minor Component ◽  
Erythroid Cell ◽  
Adherent Cell ◽  
Erythroid Cells ◽  
Gamma Chain ◽  
Single Class ◽  
Semisolid Medium ◽  
Cell Commitment

A human leukemic cell line KU 812 was recently established and described as a basophilic cell line. In the present study we show that KU 812 and two of its clones are at least bipotent: in addition to a minor component of basophils, the majority of KU 812 cells belongs to the erythroid cell lineage with a significant percentage (about 15%) of mature hemoglobinized erythroblasts. This terminal differentiation is associated with the synchronized synthesis of the main erythroid proteins, including glycophorins, spectrin beta chain, band 3, and hemoglobin. The predominant hemoglobins are adult, fetal, and Bart's hemoglobin. Adult hemoglobin represented up to 75% of all hemoglobins in the KU 812 F clone in passages containing a high number of mature erythroblasts. Transcripts of all human globin chains were present with ten times less embryonic chain messenger RNA (mRNA) than alpha-, beta- or gamma-chain mRNA. Hemin slightly increased the total hemoglobin production of the cell line, especially gamma-globin chain synthesis, but did not modify the percentage of hemoglobinized cells. Phorbol myristate acetate (PMA) had a complex effect, inducing a proportion of KU 812 cells to adhere to the plastic culture flask. The adherent cell fraction expressed a very low level of specific erythroid proteins, but their ultrastructure was consistent with immature erythroid cells. In contrast, approximately 40% of the nonadherent cells were mature erythroid cells. Cell-sorting experiments showed that this paradoxic effect of PMA is mostly due to cell selection, the more mature cells being unable to adhere. In addition, KU 812 F was found to be sensitive to erythropoietin, which slightly increased its plating efficiency range (from 0% to 50%) in semisolid medium and enhanced hemoglobin accumulation twofold. In binding experiments using 125I erythropoietin, a single class of high-affinity Epo receptors (Kd: 250 pM) was detected by binding with a density of 205 receptors per cell. The KU 812 cell line is therefore a unique model for studying cell commitment toward different hematopoietic lineages and erythroid differentiation.

scholarly journals Dissociation between p93B-myb and p75c-myb expression during the proliferation and differentiation of human myeloid cell lines

Blood ◽  
1994 ◽  
Vol 83 (7) ◽  
pp. 1778-1790 ◽  
Author(s):  
M Arsura ◽  
MM Luchetti ◽  
E Erba ◽  
J Golay ◽  
A Rambaldi ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cellular Proliferation ◽  
Myeloid Cells ◽  
Indirect Evidence ◽  
Granulocytic Differentiation ◽  
Gm Csf ◽  
Protein Levels ◽  
Proliferation And Differentiation ◽  
Myb Protein

Direct and indirect evidence strongly indicates that the proto-oncogene c-myb plays an important role in the regulation of both the proliferation and differentiation of hematopoietic cells. In addition, recent data suggest that the structurally related B-myb gene is also necessary for the proliferation of these cells. To help understand the relationship between these two related gene products during proliferation and differentiation of myeloid cells, we have studied in parallel the regulated expression of c-myb and B-myb RNAs and proteins in human myeloid cells that were either growth-arrested or induced to differentiate along different pathways. For this purpose, we have produced a polyclonal antibody directed against a fragment of the recombinant B-myb protein. We have thus been able to detect the B-myb protein in human cell lines and have found it to be a 93-kD protein localized in the nucleus. We have chosen two models to study the expression of both c-myb and B-myb mRNAs and proteins during myeloid proliferation and differentiation. One of the models was the HL-60 cell line, which can be induced to differentiate towards the monocytic pathway with either phorbol ester (phorbol myristate acetate) or vitamin D3 and towards the granulocytic pathway with either dimethyl sulfoxide or retinoic acid. In addition, we have studied another recently established human leukemic cell line, called GF-D8, which is strictly dependent on granulocyte-macrophage colony-stimulating factor (GM-CSF) for proliferation. The results show that the expression of B- myb RNA and protein closely correlates with proliferation in all experimental setups studied, whereas the c-myb protein levels do not always do so. We observed that the c-myb protein levels decreased well before the decrease of B-myb protein and of proliferation itself during differentiation toward monocytes. Such a difference was not present during granulocytic differentiation, in which c-myb levels decreased, if anything, later than those of B-myb and proliferation. Most striking was the finding that high levels of c-myb RNA and protein, but not of B- myb, were present in the GF-D8 cell line, even after growth arrest by GM-CSF deprivation. These data suggest that B-myb may function solely in the regulation of cellular proliferation, whereas c-myb has additional functions, for example, in the maintenance of an undifferentiated state.

scholarly journals KU 812: a pluripotent human cell line with spontaneous erythroid terminal maturation

Blood ◽  
1989 ◽  
Vol 73 (7) ◽  
pp. 2003-2013 ◽  
Author(s):  
M Nakazawa ◽  
MT Mitjavila ◽  
N Debili ◽  
N Casadevall ◽  
P Mayeux ◽  
...  
Keyword(s):  
Cell Line ◽  
Messenger Rna ◽  
Minor Component ◽  
Erythroid Cell ◽  
Adherent Cell ◽  
Erythroid Cells ◽  
Gamma Chain ◽  
Single Class ◽  
Semisolid Medium ◽  
Cell Commitment

Abstract A human leukemic cell line KU 812 was recently established and described as a basophilic cell line. In the present study we show that KU 812 and two of its clones are at least bipotent: in addition to a minor component of basophils, the majority of KU 812 cells belongs to the erythroid cell lineage with a significant percentage (about 15%) of mature hemoglobinized erythroblasts. This terminal differentiation is associated with the synchronized synthesis of the main erythroid proteins, including glycophorins, spectrin beta chain, band 3, and hemoglobin. The predominant hemoglobins are adult, fetal, and Bart's hemoglobin. Adult hemoglobin represented up to 75% of all hemoglobins in the KU 812 F clone in passages containing a high number of mature erythroblasts. Transcripts of all human globin chains were present with ten times less embryonic chain messenger RNA (mRNA) than alpha-, beta- or gamma-chain mRNA. Hemin slightly increased the total hemoglobin production of the cell line, especially gamma-globin chain synthesis, but did not modify the percentage of hemoglobinized cells. Phorbol myristate acetate (PMA) had a complex effect, inducing a proportion of KU 812 cells to adhere to the plastic culture flask. The adherent cell fraction expressed a very low level of specific erythroid proteins, but their ultrastructure was consistent with immature erythroid cells. In contrast, approximately 40% of the nonadherent cells were mature erythroid cells. Cell-sorting experiments showed that this paradoxic effect of PMA is mostly due to cell selection, the more mature cells being unable to adhere. In addition, KU 812 F was found to be sensitive to erythropoietin, which slightly increased its plating efficiency range (from 0% to 50%) in semisolid medium and enhanced hemoglobin accumulation twofold. In binding experiments using 125I erythropoietin, a single class of high-affinity Epo receptors (Kd: 250 pM) was detected by binding with a density of 205 receptors per cell. The KU 812 cell line is therefore a unique model for studying cell commitment toward different hematopoietic lineages and erythroid differentiation.

scholarly journals Mad4 is regulated by a transcriptional repressor complex that contains Miz-1 and c-Myc

2003 ◽  
Vol 370 (1) ◽  
pp. 291-298 ◽  
Author(s):  
Louise KIME ◽  
Stephanie C. WRIGHT
Keyword(s):  
Transcriptional Repression ◽  
Cellular Proliferation ◽  
Core Promoter ◽  
Core Promoter Region ◽  
Differentiated Cells ◽  
Chemically Induced ◽  
Proliferation And Differentiation ◽  
Repressor Complex ◽  
Undifferentiated Cells ◽  
Novel Target

Myc and Mad family proteins are central regulators of cellular proliferation and differentiation. We show that various Mad family genes have distinct patterns of expression during the chemically induced differentiation of mouse erythroleukaemia (MEL) cells, suggesting that they each serve a different function. Mad4 RNA is highly induced and persists in terminally differentiated cells, in agreement with observations in other systems. Using reporter gene assays in stably transfected MEL cells, we show that induction of Mad4 is mediated by a 49nt core promoter region. We demonstrate that the initiator element is required for Mad4 activation, and show that induction is associated with the loss from the initiator of a complex that contains Miz-1 and c-Myc. Miz-1 activates the Mad4 promoter in transient transfection assays, and this effect is antagonized by c-Myc. We therefore identify Mad4 as a novel target of transcriptional repression by c-Myc. These data suggest that the expression of Mad4 in proliferating undifferentiated cells is suppressed by the binding of a c-Myc—Miz-1 repressor complex at the initiator, and that the activation of Mad4 during differentiation results, at least in part, from a decrease in c-Myc-mediated repression.

scholarly journals Generation of a Primitive Erythroid Cell Line and Promotion of Its Growth by Basic Fibroblast Growth Factor

Blood ◽  
1998 ◽  
Vol 91 (9) ◽  
pp. 3202-3209 ◽  
Author(s):  
David Yuen ◽  
Leena Mittal ◽  
Chu-Xia Deng ◽  
Kyunghee Choi
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Cell Line ◽  
Basic Fibroblast Growth Factor ◽  
Es Cells ◽  
Embryoid Bodies ◽  
Erythroid Cell ◽  
Fibroblast Growth ◽  
Proliferation And Differentiation

An immortalized cell line representing the primitive erythroid (EryP) lineage was established from in vitro–differentiated progeny (embryoid bodies [EBs]) of embryonic stem (ES) cells using a retroviral insertional mutation, and has been termed EB-PE for embryoid body–derived primitive erythroid. Even though EB-PE cells are immortalized, they show characteristics of normal EryP cells, such as gene expression and growth factor dependency. In addition, EB-PE cells can differentiate further in culture. Investigation of growth factor requirements of EB-PE cells showed that basic fibroblast growth factor (bFGF) and erythropoietin (Epo) play unique roles in EB-PE proliferation and differentiation. While bFGF was a strong mitogen, Epo was required for both proliferation and differentiation. The unique proliferative response to bFGF coincided with upregulation of its receptor, fibroblast growth factor receptor (fgfr-1), and downregulation of erythropoietin receptor (EpoR) gene expression. Studies of primary EryP cells derived from early EBs, when tested in a colony-formation assay, also provided evidence for the mitogenic role of bFGF in concert with Epo.

Characterization of Leukemic Stem Cells in a Newly Established Acute Myeloid Leukemia (AML) Cell Line 2006-5

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 3987-3987
Author(s):  
Rui-kun Zhong ◽  
Thomas A. Lane ◽  
Edward D Ball
Keyword(s):  
Stem Cells ◽  
Cell Line ◽  
Magnetic Beads ◽  
Low Frequency ◽  
Erythroid Cell ◽  
Leukemia Stem Cells ◽  
Culture Dish ◽  
Differentiation Potential ◽  
Hla Dr ◽  
Proliferation And Differentiation

Abstract AML cell line 2006-5 was established by serial culture of an AML patients’ PBMC (M5). Compared to primary AML cells, this line expresses CD13, CD33, CD15, CD4, CD86, CD54 but lost CD34, CD64, CD123 and HLA-Dr. A group of CD13−/CD33− cells was detected consisting of 0.2% of total cells, but no significant CD34, CD90 or CD123 expression cells could be detected by FACS. After depleting of CD13+/CD33+ cells by magnetic beads, the CD13−/CD33− cells were enriched to 95%. Significant CD34+, CD90+ or CD123+ cells (0.2–0.5%) appeared in the CD13−/CD33− enriched fraction of AML cells. Leukemia colony forming (LCF) cell assay analyzed at day 14 demonstrated CD34+, CD90+ or CD123+ cells selected by magnetic beads formed typical compact large colonies of more than 1000 cells representing 40–60% of total colonies (Fig left panel). The CD13−/CD33− enriched fraction formed medium colonies with a dense center surrounded by a halo of migrating cells (80–90% of total colonies) (Fig right). Untreated 2006-5 cells formed small diffuse colonies (70–80%). Re-plating of compact colonies selected from CD34+, CD90+ or CD123+ cell LCF assays formed the above 3 type of colonies in one culture dish. Although a daughter line can be generated from a single colony formed by CD13-CD33− cells, no compact colonies could be observed in re-plating experiment. After 14 days of culture, immature erythroid cell clusters were observed in the area surrounding compact colonies suggesting that the progenitors of the colony forming leukemia cells were at the GEMM stage. Small colonies of untreated cells gradually became apoptotic and necrotic after 10 days. These results support the theory that AML is organized as a hierarchy similar to the normal hematopoietic system. CD34+, CD90+ or CD123+ cells with very low frequency of 1–2 in 1×105 2006-5 line cells demonstrated high proliferation and differentiation potential, and may represent leukemia stem cells. CD13−/CD33− cells with lower proliferation rate may belong to more differentiated leukemia progenitors without self renew potential. If additional experiments support the notion that cells forming compact colonies are leukemia stem cells, they may serve as a useful resource to test the biology of leukemia and its therapy. Figure Figure Figure Figure

PU.1 inhibits GATA-1 function and erythroid differentiation by blocking GATA-1 DNA binding

Blood ◽  
2000 ◽  
Vol 96 (8) ◽  
pp. 2641-2648 ◽  
Author(s):  
Pu Zhang ◽  
Xiaobo Zhang ◽  
Atsushi Iwama ◽  
Channing Yu ◽  
Kent A. Smith ◽  
...  
Keyword(s):  
Dna Binding ◽  
Fusion Protein ◽  
Cell Line ◽  
Protein Interactions ◽  
Messenger Rna ◽  
Erythroid Differentiation ◽  
Erythroid Cell ◽  
Protein Protein Interactions ◽  
C Terminus ◽  
N Terminus

Abstract The lineage-specific transcription factors GATA-1 and PU.1 can physically interact to inhibit each other's function, but the mechanism of repression of GATA-1 function by PU.1 has not been elucidated. Both the N terminus and the C terminus of PU.1 can physically interact with the C-terminal zinc finger of GATA-1. It is demonstrated that the PU.1 N terminus, but not the C terminus, is required for inhibiting GATA-1 function. Induced overexpression of PU.1 in K562 erythroleukemia cells blocks hemin-induced erythroid differentiation. In this system, PU.1 does not affect the expression of GATA-1 messenger RNA, protein, or nuclear localization. However, GATA-1 DNA binding decreases dramatically. By means of electrophoretic mobility shift assays with purified proteins, it is demonstrated that the N-terminal 70 amino acids of PU.1 can specifically block GATA-1 DNA binding. In addition, PU.1 had a similar effect in the G1ER cell line, in which the GATA-1 null erythroid cell line G1E has been transduced with a GATA-1–estrogen receptor fusion gene, which is directly dependent on induction of the GATA-1 fusion protein to effect erythroid maturation. Consistent with in vitro binding assays, overexpression of PU.1 blocked DNA binding of the GATA-1 fusion protein as well as GATA-1–mediated erythroid differentiation of these G1ER cells. These results demonstrate a novel mechanism by which function of a lineage-specific transcription factor is inhibited by another lineage-restricted factor through direct protein–protein interactions. These findings contribute to understanding how protein–protein interactions participate in hematopoietic differentiation and leukemogenesis.

scholarly journals Dissociation between p93B-myb and p75c-myb expression during the proliferation and differentiation of human myeloid cell lines

Blood ◽  
1994 ◽  
Vol 83 (7) ◽  
pp. 1778-1790 ◽  
Author(s):  
M Arsura ◽  
MM Luchetti ◽  
E Erba ◽  
J Golay ◽  
A Rambaldi ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cellular Proliferation ◽  
Myeloid Cells ◽  
Indirect Evidence ◽  
Granulocytic Differentiation ◽  
Gm Csf ◽  
Protein Levels ◽  
Proliferation And Differentiation ◽  
Myb Protein

Abstract Direct and indirect evidence strongly indicates that the proto-oncogene c-myb plays an important role in the regulation of both the proliferation and differentiation of hematopoietic cells. In addition, recent data suggest that the structurally related B-myb gene is also necessary for the proliferation of these cells. To help understand the relationship between these two related gene products during proliferation and differentiation of myeloid cells, we have studied in parallel the regulated expression of c-myb and B-myb RNAs and proteins in human myeloid cells that were either growth-arrested or induced to differentiate along different pathways. For this purpose, we have produced a polyclonal antibody directed against a fragment of the recombinant B-myb protein. We have thus been able to detect the B-myb protein in human cell lines and have found it to be a 93-kD protein localized in the nucleus. We have chosen two models to study the expression of both c-myb and B-myb mRNAs and proteins during myeloid proliferation and differentiation. One of the models was the HL-60 cell line, which can be induced to differentiate towards the monocytic pathway with either phorbol ester (phorbol myristate acetate) or vitamin D3 and towards the granulocytic pathway with either dimethyl sulfoxide or retinoic acid. In addition, we have studied another recently established human leukemic cell line, called GF-D8, which is strictly dependent on granulocyte-macrophage colony-stimulating factor (GM-CSF) for proliferation. The results show that the expression of B- myb RNA and protein closely correlates with proliferation in all experimental setups studied, whereas the c-myb protein levels do not always do so. We observed that the c-myb protein levels decreased well before the decrease of B-myb protein and of proliferation itself during differentiation toward monocytes. Such a difference was not present during granulocytic differentiation, in which c-myb levels decreased, if anything, later than those of B-myb and proliferation. Most striking was the finding that high levels of c-myb RNA and protein, but not of B- myb, were present in the GF-D8 cell line, even after growth arrest by GM-CSF deprivation. These data suggest that B-myb may function solely in the regulation of cellular proliferation, whereas c-myb has additional functions, for example, in the maintenance of an undifferentiated state.

Sign in / Sign up

Export Citation Format

Share Document