Acidic Leucine-Rich Nuclear Phosphoprotein 32 Family Member B (ANP32B), a Novel Caspase-3 Substrate, Exerts Anti-Apoptotic Effects on Acute Myeloid Leukemic Cells.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1337-1337
Author(s):  
Yun Yu ◽  
Shao-Ming Shen ◽  
Li-Shun Wang ◽  
Qian Zhao ◽  
Guo-Qiang Chen
Keyword(s):  
Cell Line ◽  
Nuclear Export ◽  
Caspase 3 ◽  
Expression Profiles ◽  
Leukemic Cell ◽  
Site Directed Mutagenesis ◽  
Leukemic Cells ◽  
Leukemic Cell Line ◽  
Nuclear Phosphoprotein ◽  
Acute Myeloid

Abstract The acidic leucine-rich nuclear phosphoprotein 32B (ANP32B, also called APRIL) is a member of a conserved superfamily of nuclear proteins that includes ANP32A/pp32, a factor that binds histones and inhibits their acetylation and regulates cell growth and differentiation in a tissue-specific manner. Recently, ANP32B was identified as a novel histone chaperone, and it can interact with the transcription factor KLF5, leading to transcriptional repression of a KLF5-downstream gene through stimulation of promoter region-specific histone incorporation and inhibition of histone acetylation. Additionally, ANP32B and/or ANP32A also serve as adaptor molecules linking the HuR nucleocytoplasmic shuttle protein and the nuclear export receptor CRM1 to regulate the cytoplasmic accumulation of some transcripts such as c-fos and CD83. However, its biological activity is still poorly understood. By the two-dimensional electrophoresis plus MALDI-TOF/TOF tandem mass spectrometry-based analysis of subcellular protein expression profiles, we identified ANP32B protein to become a small fragment in the cytosols of apoptotic leukemic cell line induced by NSC606985, a camptothecin analog. The ongoing immunoblot analyses confirmed that ANP32B protein was cleaved during cellular context-independent and caspase-3 activation-dependent apoptosis induced by etoposide, doxorubin and arsenic trioxide besides NSC606985. Further in vitro proteolytic experiments supported that ANP32B is a direct substrate of caspase-3, and the site-directed mutagenesis analysis identified the unclassical aspartate (AEVD163) of ANP32B sequence to be the caspase-3 targeted sites. Thus, we investigated the potential role of ANP32B in apoptosis induction. Our results showed that the suppression of ANP32B expression by siRNA in acute myeloid leukemic cell line U937 cells strongly enhances NSC606985 and etoposide-induced apoptosis. Based on these findings, this work also analyzed molecular mechanism of anti-apoptotic effect of ANP32B, and some interesting findings were confirmed.

scholarly journals Purging of acute myeloid leukemic cells by ether lipids and hyperthermia

Blood ◽  
1988 ◽  
Vol 72 (5) ◽  
pp. 1777-1783 ◽  
Author(s):  
S Okamoto ◽  
AC Olson ◽  
WE Berdel ◽  
WR Vogler
Keyword(s):  
Cell Line ◽  
Combined Treatment ◽  
Leukemic Cell ◽  
Leukemic Cells ◽  
Ether Lipids ◽  
Hematopoietic Progenitors ◽  
Leukemic Cell Line ◽  
Duration Of Treatment ◽  
Acute Myeloid

Abstract Ether lipids (EL) and hyperthermia have been shown to possess a relatively selective cytotoxicity to leukemic cells. In this study, the combined effects of EL (ET-18-OCH3, ET-16-NHCOCH3, or BM 41.440) and hyperthermia on the growth of hematopoietic progenitors, myeloid leukemic cell lines, and leukemic cells obtained from patients with acute myeloid leukemia (AML) were examined to determine if this combination resulted in a greater selective killing of leukemic cells than that achieved by either EL or heat alone. When the cells were treated simultaneously with EL (50 micrograms/mL) and hyperthermia (42 degrees C) for one hour, the killing of leukemic cell line cells was enhanced considerably. Among the three EL, however, the combination of ET-18-OCH3 and heat seemed to be the most cytotoxic to leukemic cell line cells with no effect on the growth of hematopoietic progenitors. An increase in the duration of treatment with ET-18-OCH3 to four hours with heat added during the last hour resulted in a further reduction of leukemic cell line cells while sparing 50% of hematopoietic progenitors after cryopreservation. The combined treatment with ET-18-OCH3 and heat also inhibited the growth of leukemic progenitors obtained from AML patients by 97% to 100%. These data indicate that the combined treatment with EL and hyperthermia might offer an efficient means to eliminate myeloid leukemic cells in vitro.

scholarly journals Purging of acute myeloid leukemic cells by ether lipids and hyperthermia

Blood ◽  
1988 ◽  
Vol 72 (5) ◽  
pp. 1777-1783
Author(s):  
S Okamoto ◽  
AC Olson ◽  
WE Berdel ◽  
WR Vogler
Keyword(s):  
Cell Line ◽  
Combined Treatment ◽  
Leukemic Cell ◽  
Leukemic Cells ◽  
Ether Lipids ◽  
Hematopoietic Progenitors ◽  
Leukemic Cell Line ◽  
Duration Of Treatment ◽  
Acute Myeloid

Ether lipids (EL) and hyperthermia have been shown to possess a relatively selective cytotoxicity to leukemic cells. In this study, the combined effects of EL (ET-18-OCH3, ET-16-NHCOCH3, or BM 41.440) and hyperthermia on the growth of hematopoietic progenitors, myeloid leukemic cell lines, and leukemic cells obtained from patients with acute myeloid leukemia (AML) were examined to determine if this combination resulted in a greater selective killing of leukemic cells than that achieved by either EL or heat alone. When the cells were treated simultaneously with EL (50 micrograms/mL) and hyperthermia (42 degrees C) for one hour, the killing of leukemic cell line cells was enhanced considerably. Among the three EL, however, the combination of ET-18-OCH3 and heat seemed to be the most cytotoxic to leukemic cell line cells with no effect on the growth of hematopoietic progenitors. An increase in the duration of treatment with ET-18-OCH3 to four hours with heat added during the last hour resulted in a further reduction of leukemic cell line cells while sparing 50% of hematopoietic progenitors after cryopreservation. The combined treatment with ET-18-OCH3 and heat also inhibited the growth of leukemic progenitors obtained from AML patients by 97% to 100%. These data indicate that the combined treatment with EL and hyperthermia might offer an efficient means to eliminate myeloid leukemic cells in vitro.

scholarly journals Fluorouracil Selectively Enriches Stem-like Leukemic Cells in a Leukemic Cell Line

2010 ◽  
pp. 419-427 ◽  
Author(s):  
Ling Zhang ◽  
Song Yang ◽  
Yu-Juan He ◽  
Hui-Yuan Shao ◽  
Li Wang ◽  
...  
Keyword(s):  
Cell Line ◽  
Leukemic Cell ◽  
Leukemic Cells ◽  
Leukemic Cell Line

scholarly journals Constitutive expression and role in growth regulation of interleukin-1 and multiple cytokine receptors in a biphenotypic leukemic cell line

Blood ◽  
1991 ◽  
Vol 78 (1) ◽  
pp. 94-102 ◽  
Author(s):  
A Cohen ◽  
T Grunberger ◽  
W Vanek ◽  
ID Dube ◽  
PJ Doherty ◽  
...  
Keyword(s):  
Cell Line ◽  
Growth Regulation ◽  
Interleukin 1 ◽  
Constitutive Expression ◽  
Leukemic Cell ◽  
Leukemic Cells ◽  
Cytokine Receptors ◽  
Low Density ◽  
Leukemic Cell Line ◽  
B1 Cells

A cell line (B1) was established from the bone marrow of a patient with a relapse of acute leukemia characterized by a 4;11 chromosomal translocation and biphenotypic features of early B and myeloid lineages. Analysis of the growth requirements of this cell line showed density-dependent growth and secretion of an autostimulatory growth factor, suggesting an autocrine mechanism. Several lines of evidence implicate the participation of interleukin-1 (IL-1) in the autocrine growth regulation of B1 cells. These cells constitutively express the messenger RNA (mRNA) for IL-1 and IL-1 receptor and secrete IL-1; recombinant IL-1 stimulated the growth of colonies when cells were seeded at low density, and anti-IL-1 antibodies inhibited the growth of colonies with cells seeded at higher density. B1 cells do not express detectable levels of mRNA for any of the other cytokines tested, and other cytokines failed to support the growth of B1 cells at low density. In addition, B1 cells express multiple cytokine receptor genes, including the receptors for IL-6, IL-7, tumor necrosis factor and gamma-interferon. Addition of the respective cytokines to the B1 cells resulted in inhibition of the growth of leukemic cells in vitro. The multiplicity of growth-inhibitory cytokine receptors on this leukemic cell line might be due to its biphenotypic lineage and may suggest new therapeutic possibilities in controlling leukemic cell proliferation.

scholarly journals LacZ staining in paraffin-embedded tissue sections.

1996 ◽  
Vol 44 (11) ◽  
pp. 1323-1329 ◽  
Author(s):  
P J Hendrikx ◽  
J Vermeulen ◽  
A Hagenbeek ◽  
M Vermey ◽  
A C Martens
Keyword(s):  
Cell Line ◽  
Leukemic Cell ◽  
Promyelocytic Leukemia ◽  
Leukemic Cells ◽  
Leukemic Cell Line ◽  
Paraffin Sections ◽  
Tissue Sections ◽  
Lacz Staining ◽  
Catalyzed Reporter Deposition ◽  
First Time

Femora and tibiae of rats carrying leukemia from a LacZ-marked acute promyelocytic leukemia-derived leukemic cell line (LT12NL15) were decalcified using EDTA and routinely embedded in paraffin. Sections were used to develop for the first time an immunostaining method for LacZ, employing catalyzed reporter deposition (CARD) based on the deposition of biotinylated tyramine. This method was used to study homing and adhesion of leukemic cells.

scholarly journals Constitutive expression and role in growth regulation of interleukin-1 and multiple cytokine receptors in a biphenotypic leukemic cell line

Blood ◽  
1991 ◽  
Vol 78 (1) ◽  
pp. 94-102 ◽  
Author(s):  
A Cohen ◽  
T Grunberger ◽  
W Vanek ◽  
ID Dube ◽  
PJ Doherty ◽  
...  
Keyword(s):  
Cell Line ◽  
Growth Regulation ◽  
Interleukin 1 ◽  
Constitutive Expression ◽  
Leukemic Cell ◽  
Leukemic Cells ◽  
Cytokine Receptors ◽  
Low Density ◽  
Leukemic Cell Line ◽  
B1 Cells

Abstract A cell line (B1) was established from the bone marrow of a patient with a relapse of acute leukemia characterized by a 4;11 chromosomal translocation and biphenotypic features of early B and myeloid lineages. Analysis of the growth requirements of this cell line showed density-dependent growth and secretion of an autostimulatory growth factor, suggesting an autocrine mechanism. Several lines of evidence implicate the participation of interleukin-1 (IL-1) in the autocrine growth regulation of B1 cells. These cells constitutively express the messenger RNA (mRNA) for IL-1 and IL-1 receptor and secrete IL-1; recombinant IL-1 stimulated the growth of colonies when cells were seeded at low density, and anti-IL-1 antibodies inhibited the growth of colonies with cells seeded at higher density. B1 cells do not express detectable levels of mRNA for any of the other cytokines tested, and other cytokines failed to support the growth of B1 cells at low density. In addition, B1 cells express multiple cytokine receptor genes, including the receptors for IL-6, IL-7, tumor necrosis factor and gamma-interferon. Addition of the respective cytokines to the B1 cells resulted in inhibition of the growth of leukemic cells in vitro. The multiplicity of growth-inhibitory cytokine receptors on this leukemic cell line might be due to its biphenotypic lineage and may suggest new therapeutic possibilities in controlling leukemic cell proliferation.

IκB Kinase Overcomes PI3K to Control FoxO3a Subcellular Localization in Acute Myeloid Leukemia.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 478-478 ◽  
Author(s):  
Nicolas Chapuis ◽  
Sophie Park ◽  
Laurent Leotoing ◽  
Jerome Tamburini ◽  
Frederique Verdier ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Subcellular Localization ◽  
Cell Line ◽  
Nuclear Export ◽  
Leukemic Cell ◽  
Pi3k Activity ◽  
Iκb Kinase ◽  
Blast Cells ◽  
Fas L ◽  
Acute Myeloid

Abstract Abstract 478 The prognosis of Acute Myeloid Leukaemia (AML) remains globally poor. Aberrant activation of signalling pathways is frequently found in AML and leads to uncontrolled cell growth and survival. Effective targeting of these pathways with new therapeutics may result in suppression of leukemic cell proliferation and survival. In AML, constitutive PI3K activity, mainly due to the expression of the class IA PI3K p110δ isoform, is detected in 50% of patients at diagnosis (PI3K+ AML samples) (Sujobert, Bardet et al., Blood 2005; Tamburini, Elie et al., Blood 2007). The PI3K/Akt axis represents therefore an attractive therapeutic target in this disease. Accordingly, inhibition of PI3K activity with IC87114, a specific p110δ inhibitor decreases AML blast cells proliferation (Sujobert, Bardet et al., Blood 2005). However, IC87114 failed to induce significant apoptosis (Park, Chapuis et al., Leukemia 2008). The PI3K/Akt network controls different targets implicated in the regulation of cell survival. Among them, the FoxO transcription factors which include FoxO1, FoxO3a, FoxO4 and FoxO6 up regulate the expression of target genes involved in apoptosis, such Fas-L and Bim. Akt, by phosphorylating FoxO proteins at three conserved sites (T32, S253 and S315 on FoxO3a), negatively regulates their transcriptional activity by inducing their nuclear export. Given that IC87114 did not induce apoptosis, we hypothesized that FoxO proteins may escape to PI3K/Akt control in AML cells. For that purpose, we investigated their mechanisms of regulation in both primary AML cells and a MV4-11 human leukemic cell line engineered to stably overexpress a FoxO3awt-GPF fusion protein introduced by lentiviral infection. We focused especially on FoxO3a as we found, using real-time RT-PCR and WB analysis, that it was the only FoxO protein constantly expressed in primary AML cells. First, we examined the relationship between FoxO3a subcellular localization using immunofluorescence and PI3K activity in primary AML cells. As expected in PI3K+ AML cells (n=6), FoxO3a was phosphorylated on T32 and S253 and constantly localized in the cytoplasm. However, although Akt S473 and FoxO3a T32 and S253 phosphorylations were fully inhibited by IC87114, FoxO3a did not translocate into the nucleus of blast cells. Accordingly, Fas-L and Bim expression tested by qRT-PCR were not induced by IC87114. Identical results were obtained from the same analysis of the MV4-11FoxO3awt-GFP cell line. Furthermore, in PI3K- AML cells (n=5), neither Akt S473 nor FoxO3a T32 and S253 phosphorylations were detected but FoxO3a was still confined in the cytoplasm of these cells. Altogether, these results clearly demonstrate that an Akt-independent mechanism contributes to the nuclear exclusion of FoxO3a in AML cells. In addition to Akt, the IκB Kinase (IKK) may regulate FoxO3a subcellular localization by phosphorylation on S644 in AML, as it has previously been shown in breast cancer (Hu, Lee et al., Cell 2004). As the oncogenic IκB Kinase is known to be activated in almost all AML samples (Guzman, Neering et al., Blood 2001), we investigated whether IKK could maintain FoxO3a in the cytoplasm of AML cells. Interestingly, we found that inhibition of IKK with a specific IKKγ/NEMO-antagonistic peptide (Agou, Courtois et al., J Biol Chem 2004) induced a strong nuclear translocation of FoxO3a and an efficient apoptosis in both primary AML blast cells (n=5) and MV4-11FoxO3awt-GFP cell line. To confirm this result, MV4-11 cells were infected with a lentivirus expressing a FoxO3aS644A-GFP mutant protein in which the IKK phosphorylation is abrogated. We found that the FoxO3aS644A-GFP mutant protein primarily localized within the nucleus of MV4-11 cells and induced a decrease of cell proliferation and a moderate pro-apoptotic effect. We thus conclude that IKK, by phosphorylating FoxO3a on S644, leads to the constant nuclear export of FoxO3a in AML cells. Consequently, the therapeutical potential value of IKK targeted inhibition in AML could be due not only to the inhibition of NF-κB activity but also to the FoxO3a nuclear import. Moreover, the PI3K-independent FoxO3a regulation probably contributes to the low potential of specific PI3K inhibitors to induce apoptosis in AML cells. Disclosures: No relevant conflicts of interest to declare.

Sign in / Sign up

Export Citation Format

Share Document