Overexpression of the human manganese superoxide dismutase (MnSOD) transgene in subclones of murine hematopoietic progenitor cell line 32D cl 3 decreases irradiation-induced apoptosis but does not alter G2/M or G1/S phase cell cycle arrest

Abstract We investigated the involvement of NF-κB/Rel transcription factors that reportedly can inhibit apoptosis in various cell types in the antiapoptotic mechanism of the cytoprotectant amifostine. In the nontumorigenic murine myeloid progenitor 32D cells incubated with amifostine, we detected a reduction of the IκB cytoplasmic levels by Western blotting and a raising of nuclear NF-κB/Rel complexes by electrophoretic mobility shift assay. Amifostine inhibited by more than 30% the growth factor deprivation-induced apoptosis, whereas its effect failed when we blocked the NF-κB/Rel activity with an NF-κB/Rel-binding phosphorothioate decoy oligodeoxynucleotide. In human cord blood CD34+ cells, the NF-κB/Rel p65 subunit was detectable (using immunofluorescence analysis) mainly in the cytoplasm in the absence of amifostine, whereas its presence was appreciable in the nuclei of cells incubated with the cytoprotectant. In 4 CD34+ samples incubated for 3 days in cytokine-deficient conditions, cell apoptosis was reduced by more than 30% in the presence of amifostine (or amifostine plus a control oligo); the effect of amifostine was abolished in cultures with the decoy oligo. These findings indicate that the inhibition of hematopoietic progenitor cell apoptosis by amifostine requires the induction of NF-κB/Rel factors and that the latter can therefore exert an antiapoptotic activity in the hematopoietic progenitor cell compartment. Furthermore, the identification of this specific mechanism underlying the survival-promoting activity of amifostine lends support to the possible use of this agent in apoptosis-related pathologies, such as myelodysplasias.

Download Full-text

Mapping gene expression patterns during myeloid differentiation using the EML hematopoietic progenitor cell line

Experimental Hematology ◽

10.1016/s0301-472x(02)00817-2 ◽

2002 ◽

Vol 30 (7) ◽

pp. 649-658 ◽

Cited By ~ 7

Author(s):

Yang Du ◽

Janee L Campbell ◽

Demet Nalbant ◽

Hyewon Youn ◽

Ann C.Hughes Bass ◽

...

Keyword(s):

Gene Expression ◽

Cell Line ◽

Progenitor Cell ◽

Expression Patterns ◽

Hematopoietic Progenitor Cell ◽

Myeloid Differentiation ◽

Gene Expression Patterns ◽

Hematopoietic Progenitor ◽

Mapping Gene ◽

Progenitor Cell Line

Download Full-text

Optimal Proliferation of a Hematopoietic Progenitor Cell Line Requires Either Costimulation With Stem Cell Factor or Increase of Receptor Expression That Can Be Replaced by Overexpression of Bcl-2

Blood ◽

10.1182/blood.v93.8.2569 ◽

1999 ◽

Vol 93 (8) ◽

pp. 2569-2577 ◽

Cited By ~ 18

Author(s):

Huei-Mei Huang ◽

Jian-Chiuan Li ◽

Yueh-Chun Hsieh ◽

Hsin-Fang Yang-Yen ◽

Jeffrey Jong-Young Yen

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Cell Line ◽

Progenitor Cells ◽

Stem Cell Factor ◽

Progenitor Cell ◽

Stem Cell Differentiation ◽

Hematopoietic Progenitor Cell ◽

Hematopoietic Progenitor ◽

Progenitor Cell Line

Abstract In vitro proliferation of hematopoietic stem cells requires costimulation by multiple regulatory factors whereas expansion of lineage-committed progenitor cells generated by stem cells usually requires only a single factor. The distinct requirement of factors for proliferation coincides with the differential temporal expression of the subunits of cytokine receptors during early stem cell differentiation. In this study, we explored the underlying mechanism of the requirement of costimulation in a hematopoietic progenitor cell line TF-1. We found that granulocyte-macrophage colony-stimulating factor (GM-CSF) optimally activated proliferation of TF-1 cells regardless of the presence or absence of stem cell factor (SCF). However, interleukin-5 (IL-5) alone sustained survival of TF-1 cells and required costimulation of SCF for optimal proliferation. The synergistic effect of SCF was partly due to its anti-apoptosis activity. Overexpression of the IL-5 receptor  subunit (IL5R) in TF-1 cells by genetic selection or retroviral infection also resumed optimal proliferation due to correction of the defect in apoptosis suppression. Exogenous expression of an oncogenic anti-apoptosis protein, Bcl-2, conferred on TF-1 cells an IL-5–dependent phenotype. In summary, our data suggested SCF costimulation is only necessary when the expression level of IL5R is low and apoptosis suppression is defective in the signal transduction of IL-5. Expression of Bcl-2 proteins released the growth restriction of the progenitor cells and may be implicated in leukemia formation.

Download Full-text

Optimal Proliferation of a Hematopoietic Progenitor Cell Line Requires Either Costimulation With Stem Cell Factor or Increase of Receptor Expression That Can Be Replaced by Overexpression of Bcl-2

Blood ◽

10.1182/blood.v93.8.2569.408k08_2569_2577 ◽

1999 ◽

Vol 93 (8) ◽

pp. 2569-2577 ◽

Cited By ~ 1

Author(s):

Huei-Mei Huang ◽

Jian-Chiuan Li ◽

Yueh-Chun Hsieh ◽

Hsin-Fang Yang-Yen ◽

Jeffrey Jong-Young Yen

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Cell Line ◽

Progenitor Cells ◽

Stem Cell Factor ◽

Progenitor Cell ◽

Stem Cell Differentiation ◽

Hematopoietic Progenitor Cell ◽

Hematopoietic Progenitor ◽

Progenitor Cell Line

In vitro proliferation of hematopoietic stem cells requires costimulation by multiple regulatory factors whereas expansion of lineage-committed progenitor cells generated by stem cells usually requires only a single factor. The distinct requirement of factors for proliferation coincides with the differential temporal expression of the subunits of cytokine receptors during early stem cell differentiation. In this study, we explored the underlying mechanism of the requirement of costimulation in a hematopoietic progenitor cell line TF-1. We found that granulocyte-macrophage colony-stimulating factor (GM-CSF) optimally activated proliferation of TF-1 cells regardless of the presence or absence of stem cell factor (SCF). However, interleukin-5 (IL-5) alone sustained survival of TF-1 cells and required costimulation of SCF for optimal proliferation. The synergistic effect of SCF was partly due to its anti-apoptosis activity. Overexpression of the IL-5 receptor  subunit (IL5R) in TF-1 cells by genetic selection or retroviral infection also resumed optimal proliferation due to correction of the defect in apoptosis suppression. Exogenous expression of an oncogenic anti-apoptosis protein, Bcl-2, conferred on TF-1 cells an IL-5–dependent phenotype. In summary, our data suggested SCF costimulation is only necessary when the expression level of IL5R is low and apoptosis suppression is defective in the signal transduction of IL-5. Expression of Bcl-2 proteins released the growth restriction of the progenitor cells and may be implicated in leukemia formation.

Download Full-text

Amifostine Inhibits Hematopoietic Progenitor Cell Apoptosis by Activating NF-κB/Rel Transcription Factors

Blood ◽

10.1182/blood.v94.12.4060.424k23_4060_4066 ◽

1999 ◽

Vol 94 (12) ◽

pp. 4060-4066 ◽

Cited By ~ 2

Author(s):

Maria Fiammetta Romano ◽

Annalisa Lamberti ◽

Rita Bisogni ◽

Corrado Garbi ◽

Antonio M. Pagnano ◽

...

Keyword(s):

Transcription Factors ◽

Cell Apoptosis ◽

Progenitor Cell ◽

Cell Types ◽

Hematopoietic Progenitor Cell ◽

Hematopoietic Progenitor ◽

Mobility Shift ◽

Human Cord Blood ◽

Mobility Shift Assay ◽

Induced Apoptosis

We investigated the involvement of NF-κB/Rel transcription factors that reportedly can inhibit apoptosis in various cell types in the antiapoptotic mechanism of the cytoprotectant amifostine. In the nontumorigenic murine myeloid progenitor 32D cells incubated with amifostine, we detected a reduction of the IκB cytoplasmic levels by Western blotting and a raising of nuclear NF-κB/Rel complexes by electrophoretic mobility shift assay. Amifostine inhibited by more than 30% the growth factor deprivation-induced apoptosis, whereas its effect failed when we blocked the NF-κB/Rel activity with an NF-κB/Rel-binding phosphorothioate decoy oligodeoxynucleotide. In human cord blood CD34+ cells, the NF-κB/Rel p65 subunit was detectable (using immunofluorescence analysis) mainly in the cytoplasm in the absence of amifostine, whereas its presence was appreciable in the nuclei of cells incubated with the cytoprotectant. In 4 CD34+ samples incubated for 3 days in cytokine-deficient conditions, cell apoptosis was reduced by more than 30% in the presence of amifostine (or amifostine plus a control oligo); the effect of amifostine was abolished in cultures with the decoy oligo. These findings indicate that the inhibition of hematopoietic progenitor cell apoptosis by amifostine requires the induction of NF-κB/Rel factors and that the latter can therefore exert an antiapoptotic activity in the hematopoietic progenitor cell compartment. Furthermore, the identification of this specific mechanism underlying the survival-promoting activity of amifostine lends support to the possible use of this agent in apoptosis-related pathologies, such as myelodysplasias.

Download Full-text

Erythropoietin increases the radioresistance of a clonal hematopoietic progenitor cell line expressing a transgene for the erythropoietin receptor

Stem Cells ◽

10.1002/stem.5530120506 ◽

1994 ◽

Vol 12 (5) ◽

pp. 506-513 ◽

Cited By ~ 10

Author(s):

Maria Alessandra Santucci ◽

Jacalyn H. Pierce ◽

Stella Zannini ◽

Alessandra Fortuna ◽

Giovanni Frezza ◽

...

Keyword(s):

Cell Line ◽

Progenitor Cell ◽

Erythropoietin Receptor ◽

Hematopoietic Progenitor Cell ◽

Hematopoietic Progenitor ◽

Progenitor Cell Line

Download Full-text

Functional Interaction of c-Kit Receptor and Interleukin-3 Receptor.

Blood ◽

10.1182/blood.v110.11.2228.2228 ◽

2007 ◽

Vol 110 (11) ◽

pp. 2228-2228

Author(s):

Zhijia Ye ◽

Sherman M. Weissman

Keyword(s):

Cell Line ◽

Progenitor Cell ◽

Pi3 Kinase ◽

Hematopoietic Progenitor Cell ◽

Hematopoietic Progenitor ◽

Interleukin 3 ◽

Gm Csf ◽

Β Chain ◽

Progenitor Cell Line ◽

Negative Population

Abstract Hematopoiesis is regulated by a number of growth factors and cytokines, among which stem cell factor (SCF) plays a critical role for normal hematopoiesis. An important feature of SCF is its strong capacity to synergize with the cytokines including interleukin-3(IL-3), granulocyte-marcrophage colony-stimulation factor (GM-CSF) and erythropoietin to regulate proliferation and differentiations of hematopoietic progenitor cells. The mouse hematopoietic progenitor cell line EML contains two populations of lineage negative cells, distinguished by the expression or absence of CD34. The two sub-populations express similar levels of the SCF receptor c-kit, but only the CD34+ population replicates in the presence of SCF alone, while the CD34 negative population replicates in the presence of IL3 but this replication is synergistically stimulated by the addition of SCF. Our previous studies indicated that synergistic proliferation of the mouse hematopoietic progenitor cell line EML induced by SCF in combination with IL-3 was probably mediated by the synergistic increases in tyrosine phosphorylation of c-Kit and β-chain of IL-3 receptor, which is shared with GM-CSF receptor and interleukin-5 receptor. Conversely, the decreased response of c-kit to SCF alone in the CD34 negative population may be a result of the increased expression of IL3 receptor in these cells. Trans-phosphorylation between c-Kit and the β-chain of IL-3 receptor was also observed in EML cells. In studies of the molecular mechanism behind the functional interactions between SCF and IL-3 we found that c-Kit and IL-3 receptor β-chain form a complex in EML cells. Antibody Ab-1 specific against the Ig-like domain 4 in the extracellular region of c-Kit blocked not only cell proliferation induced by SCF but also synergistic proliferation induced by SCF plus IL-3. Consistent with this finding, Ab-1 inhibited phosphorylation of c-Kit induced by SCF or IL-3, and synergistic phosphorylation of c-kit induced by SCF plus IL-3 respectively. SCF and IL-3 in combination synergistically activated the protein kinases Akt and Erks, the downstream mediators of c-Kit and IL-3R. Phosphorylation of Erks is independent on the phosphatidylinositol 3-kinase (PI3-kinase) pathway in EML cells. These data suggested the possibility that the synergistic growth of EML cells induced by SCF in combination with IL-3 was mediated by multiple mechanisms that could include trans- and synergistic phosphorylation of c-Kit and the β-chain of IL-3 receptor, PI3-kinase dependent synergistic phosphorylation of Akt and PI3-kinase independent synergistic phosphorylation of Erks. The trans- and synergistic phosphorylation of c-Kit and β-chain of IL-3 receptor was mediated by physical interaction of two receptors.

Download Full-text