Retinoic acid-induced granulocytic differentiation of HL-60 myeloid leukemia cells is mediated directly through the retinoic acid receptor (RAR-alpha)

1990 ◽  
Vol 10 (5) ◽  
pp. 2154-2163
Author(s):  
S J Collins ◽  
K A Robertson ◽  
L Mueller
Keyword(s):  
Retinoic Acid ◽  
Myeloid Leukemia ◽  
Retinoic Acid Receptor ◽  
Leukemia Cell ◽  
Single Copy ◽  
Leukemia Cell Line ◽  
Leukemia Cells ◽  
Granulocytic Differentiation ◽  
Myeloid Leukemias ◽  
Promyelocytic Leukemia Cell Line

Retinoic acid (RA) induces terminal granulocytic differentiation of the HL-60 promyelocytic leukemia cell line as well as certain other human myeloid leukemias. Specific RA receptors that are members of the steroid-thyroid hormone superfamily of nuclear transcription factors have recently been identified. We developed an HL-60 subclone that was relatively resistant to RA-induced differentiation. Specific nuclear RA receptors in this RA-resistant subclone had a decreased affinity for RA and exhibited a lower molecular weight compared with nuclear RA receptors from the RA-sensitive parental HL-60 cells. Retroviral vector-mediated transduction of a single copy of the RA receptor (RAR-alpha) into this RA-resistant HL-60 subclone restored the sensitivity of these cells to RA. These observations indicate that RAR-alpha plays a critical and central role in mediating RA-induced terminal differentiation of HL-60 leukemia cells.

scholarly journals Retinoic acid-induced granulocytic differentiation of HL-60 myeloid leukemia cells is mediated directly through the retinoic acid receptor (RAR-alpha).

1990 ◽  
Vol 10 (5) ◽  
pp. 2154-2163 ◽  
Author(s):  
S J Collins ◽  
K A Robertson ◽  
L Mueller
Keyword(s):  
Retinoic Acid ◽  
Myeloid Leukemia ◽  
Retinoic Acid Receptor ◽  
Leukemia Cell ◽  
Single Copy ◽  
Leukemia Cell Line ◽  
Leukemia Cells ◽  
Granulocytic Differentiation ◽  
Myeloid Leukemias ◽  
Promyelocytic Leukemia Cell Line

Retinoic acid (RA) induces terminal granulocytic differentiation of the HL-60 promyelocytic leukemia cell line as well as certain other human myeloid leukemias. Specific RA receptors that are members of the steroid-thyroid hormone superfamily of nuclear transcription factors have recently been identified. We developed an HL-60 subclone that was relatively resistant to RA-induced differentiation. Specific nuclear RA receptors in this RA-resistant subclone had a decreased affinity for RA and exhibited a lower molecular weight compared with nuclear RA receptors from the RA-sensitive parental HL-60 cells. Retroviral vector-mediated transduction of a single copy of the RA receptor (RAR-alpha) into this RA-resistant HL-60 subclone restored the sensitivity of these cells to RA. These observations indicate that RAR-alpha plays a critical and central role in mediating RA-induced terminal differentiation of HL-60 leukemia cells.

scholarly journals Retinoic acid receptors in myeloid leukemia: characterization of receptors in retinoic acid-resistant K-562 cells

Blood ◽  
1991 ◽  
Vol 77 (2) ◽  
pp. 340-347 ◽  
Author(s):  
KA Robertson ◽  
L Mueller ◽  
SJ Collins
Keyword(s):  
Retinoic Acid ◽  
Myeloid Leukemia ◽  
Retinoic Acid Receptor ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Leukemia Cells ◽  
Approximate Molecular Weight ◽  
K 562 Cells ◽  
Hemoglobin Production

Although mRNA for the retinoic acid receptor alpha (RAR-alpha) is expressed in many different myeloid leukemias, most of these leukemia cells exhibit little if any phenotypic response when exposed to retinoic acid (RA). To determine whether such RA resistance is related to altered RA receptor structure or function, we performed a detailed analysis of nuclear RA receptors in RA-resistant K-562 cells. These cells exhibit RA receptors of the same approximate molecular weight and similar kd as those exhibited by the RA-sensitive HL-60 leukemia cell line, but the number of RA receptors in the RA-resistant K-562 cells (80 per cell) is significantly lower than that exhibited by RA- sensitive HL-60 cells (550 per cell). Retroviral-mediated transduction of RAR-alpha cDNA into K-562 significantly increased the number of RA receptors to 2,000 per cell. These RAR-alpha-transduced K-562 cells, when incubated with RA, exhibit diminished cell proliferation associated with decreased c-myc expression and an accumulation of cells in G0/G1. In addition, these RA-treated cells exhibit downregulation of the CD15 surface antigen and a slight increase in hemoglobin production but manifest no other evidence of significant erythroid, megakaryocytic, or myeloid differentiation. These results indicate that an elevated number of nuclear RA receptors can be involved in altering proliferation but not necessarily the differentiation of certain RA- treated myeloid leukemia cells.

scholarly journals Retinoic acid receptors in myeloid leukemia: characterization of receptors in retinoic acid-resistant K-562 cells

Blood ◽  
1991 ◽  
Vol 77 (2) ◽  
pp. 340-347 ◽  
Author(s):  
KA Robertson ◽  
L Mueller ◽  
SJ Collins
Keyword(s):  
Retinoic Acid ◽  
Myeloid Leukemia ◽  
Retinoic Acid Receptor ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Leukemia Cells ◽  
Approximate Molecular Weight ◽  
K 562 Cells ◽  
Hemoglobin Production

Abstract Although mRNA for the retinoic acid receptor alpha (RAR-alpha) is expressed in many different myeloid leukemias, most of these leukemia cells exhibit little if any phenotypic response when exposed to retinoic acid (RA). To determine whether such RA resistance is related to altered RA receptor structure or function, we performed a detailed analysis of nuclear RA receptors in RA-resistant K-562 cells. These cells exhibit RA receptors of the same approximate molecular weight and similar kd as those exhibited by the RA-sensitive HL-60 leukemia cell line, but the number of RA receptors in the RA-resistant K-562 cells (80 per cell) is significantly lower than that exhibited by RA- sensitive HL-60 cells (550 per cell). Retroviral-mediated transduction of RAR-alpha cDNA into K-562 significantly increased the number of RA receptors to 2,000 per cell. These RAR-alpha-transduced K-562 cells, when incubated with RA, exhibit diminished cell proliferation associated with decreased c-myc expression and an accumulation of cells in G0/G1. In addition, these RA-treated cells exhibit downregulation of the CD15 surface antigen and a slight increase in hemoglobin production but manifest no other evidence of significant erythroid, megakaryocytic, or myeloid differentiation. These results indicate that an elevated number of nuclear RA receptors can be involved in altering proliferation but not necessarily the differentiation of certain RA- treated myeloid leukemia cells.

scholarly journals Terminal differentiation of human promyelocytic leukemic cells in primary culture in response to retinoic acid

Blood ◽  
1981 ◽  
Vol 57 (6) ◽  
pp. 1000-1004 ◽  
Author(s):  
TR Breitman ◽  
SJ Collins ◽  
BR Keene
Keyword(s):  
Retinoic Acid ◽  
Acute Promyelocytic Leukemia ◽  
Leukemia Cell ◽  
Promyelocytic Leukemia ◽  
Leukemic Cells ◽  
Leukemia Cell Line ◽  
Granulocytic Differentiation ◽  
Myelocytic Leukemia ◽  
Functional Maturation ◽  
Promyelocytic Leukemia Cell Line

The recent finding that retinoic acid induces terminal granulocytic differentiation of the human promyelocytic leukemia cell line, HL-60, prompted an investigation of the sensitivity to this inducer of human myelocytic leukemia cells in primary suspension culture. Of the 21 leukemic specimens, only cells from the two patients with acute promyelocytic leukemia differentiated in response to retinoic acid. After an incubation period of 5--7 days in 1 microM retinoic acid, the cells from these two patients showed extensive morphological and functional maturation. Thus, because it appears that retinoic acid specifically induces granulocytic differentiation of leukemic promyelocytes, this compound may have therapeutic utility in the treatment of acute promyelocytic leukemia.

scholarly journals Multiple members of the retinoic acid receptor family are capable of mediating the granulocytic differentiation of HL-60 cells.

1992 ◽  
Vol 12 (9) ◽  
pp. 3743-3749 ◽  
Author(s):  
K A Robertson ◽  
B Emami ◽  
L Mueller ◽  
S J Collins
Keyword(s):  
Retinoic Acid ◽  
Retinoic Acid Receptor ◽  
Biological Effects ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Critical Threshold ◽  
Granulocytic Differentiation ◽  
Specific Receptors ◽  
The Individual ◽  
Receptor Family

The complex and diverse biological effects of retinoic acid (RA) are mediated through specific receptors that are members of the steroid hormone family of nuclear transcription factors. The RA receptor family consists of multiple structurally distinct RA receptors, which diverge primarily at the NH2-terminal domain. The evolutionary conservation of this divergent region in individual RA receptors among different species together with their tissue-specific patterns of expression suggest that the biological function and activity of the individual RA receptors may be confined to specific tissues. To test this hypothesis in hematopoietic cells, we used retrovirus-mediated gene transduction to introduce the RA receptors RAR-alpha, RAR-beta, and RAR-gamma as well as RXR-alpha into a mutant subclone of the HL-60 promyelocytic leukemia cell line (designated HL-60R) that is relatively resistant to RA-induced granulocytic differentiation. We found that each of these structurally distinct RA receptors could restore sensitivity of the HL-60R cells to RA. A critical threshold number of transduced receptors per cell appears to be necessary to restore this functional activity. Thus, the capability to mediate granulocytic differentiation of HL-60 cells is shared among distinctly different RA receptors.

Multiple members of the retinoic acid receptor family are capable of mediating the granulocytic differentiation of HL-60 cells

1992 ◽  
Vol 12 (9) ◽  
pp. 3743-3749
Author(s):  
K A Robertson ◽  
B Emami ◽  
L Mueller ◽  
S J Collins
Keyword(s):  
Retinoic Acid ◽  
Retinoic Acid Receptor ◽  
Biological Effects ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Critical Threshold ◽  
Granulocytic Differentiation ◽  
Specific Receptors ◽  
The Individual ◽  
Receptor Family

The complex and diverse biological effects of retinoic acid (RA) are mediated through specific receptors that are members of the steroid hormone family of nuclear transcription factors. The RA receptor family consists of multiple structurally distinct RA receptors, which diverge primarily at the NH2-terminal domain. The evolutionary conservation of this divergent region in individual RA receptors among different species together with their tissue-specific patterns of expression suggest that the biological function and activity of the individual RA receptors may be confined to specific tissues. To test this hypothesis in hematopoietic cells, we used retrovirus-mediated gene transduction to introduce the RA receptors RAR-alpha, RAR-beta, and RAR-gamma as well as RXR-alpha into a mutant subclone of the HL-60 promyelocytic leukemia cell line (designated HL-60R) that is relatively resistant to RA-induced granulocytic differentiation. We found that each of these structurally distinct RA receptors could restore sensitivity of the HL-60R cells to RA. A critical threshold number of transduced receptors per cell appears to be necessary to restore this functional activity. Thus, the capability to mediate granulocytic differentiation of HL-60 cells is shared among distinctly different RA receptors.

Redox Regulation of Retinoic Acid Receptor Function in Myeloid Leukemia Cell Differentiation and Apoptosis.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 2483-2483
Author(s):  
Kent A. Robertson ◽  
Meihua Luo ◽  
Ashley Chastain ◽  
Scott Colvin ◽  
April Reed ◽  
...  
Keyword(s):  
Cell Death ◽  
Retinoic Acid ◽  
Programmed Cell Death ◽  
Growth Inhibition ◽  
Myeloid Leukemia ◽  
Redox Regulation ◽  
Retinoic Acid Receptor ◽  
Leukemia Cell ◽  
Granulocytic Differentiation ◽  
Acid Receptor

Abstract Ape1/ref-1 is a multifunctional base excision DNA repair protein that is involved in the repair of abasic sites in DNA. However, it also has a distinct role in the redox regulation of a variety of cellular proteins, such as Fos, Jun, p53, NFkB, PAX, HIF-1a, HLF, and others. Ape-1/ref-1 maintains these proteins in a reduced state thereby facilitating their DNA binding and transcriptional activation capability. HL-60 cells are known to respond to retinoic acid (RA) with terminal granulocytic differentiation and apoptosis, which is mediated through the RA receptors. Previous experiments suggested that Ape1/ref-1 expression is related to apoptosis. To further define this relationship, we used retroviral gene transduction to over-express HA-tagged Ape1/ref-1 in HL-60 myeloid leukemia cells. We observed that the RA-induced growth inhibition of HL-60 cells over-expressing Ape1/ref-1 was significantly enhanced compared to wild type HL-60 cells. To determine if the growth inhibition was related to enhanced programmed cell death and differentiation, we treated Ape1/ref-1 transduced and vector-only (LXSN) transduced HL-60 cells with RA and evaluated the expression of Ape1/ref-1 and the development of apoptosis and markers of differentiation. Results: 1) RA induced expression of the retroviral Ape1/ref-1 construct as determined by Western blot resulting in a higher (ie retroviral + endogenous Ape1/ref-1) overall expression of Ape1/ref-1 compared to control cells; 2) analysis of RA-treated cells for apoptosis by propidium iodide, TUNEL, and Annexin V staining as well as morphology, unexpectedly demonstrated enhanced programmed cell death in cells expressing the transduced Ape1/ref-1; 3) Ape-1 over-expression enhanced the retinoid differentiation response by morphology and expression of CD11b. Additional mobility shift experiments demonstrated the redox dependence of retinoic acid receptor binding to retinoid response elements mediated by Ape-1/ref-1. In conclusion, our data supports the contention that Ape1/ref-1 expression may be important for mediating RA-induced myeloid differentiation and programmed cell death.

Ape-1/ref-1 Mediated Redox Regulation of Retinoic Acid Receptor Transcription Function in Myeloid Leukemia Cell Differentiation and Apoptosis.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1924-1924
Author(s):  
Kent A. Robertson ◽  
Edwin S. Colvin ◽  
Meihua Luo ◽  
April Reed ◽  
Mark R. Kelley
Keyword(s):  
Retinoic Acid ◽  
Myeloid Leukemia ◽  
Redox Regulation ◽  
Retinoic Acid Receptor ◽  
Leukemia Cell ◽  
Luciferase Reporter ◽  
Luciferase Expression ◽  
Granulocytic Differentiation ◽  
Acid Receptor ◽  
Retinoic Acid Response Element

Abstract Ape1/ref-1is a multifunctional base excision DNA repair protein that is involved in the repair of abasic sites in DNA. However, it also has a distinct role in the redox regulation of a variety of cellular proteins, such as Fos, Jun, p53, NFkB, PAX, HIF-1a, HLF, and others. Ape-1/ref-1 maintains these proteins in a reduced state thereby facilitating their DNA binding and transcriptional activation capability. HL-60 cells are known to respond to retinoic acid (RA) with terminal granulocytic differentiation and apoptosis, which is mediated through the RA receptors. Previous experiments suggested that Ape1/ref-1 expression is related to apoptosis. To further define this relationship, we used retroviral gene transduction to over-express HA-tagged Ape1/ref-1 in HL-60 myeloid leukemia cells. We observed that the RA-induced growth inhibition, apoptosis, and differentiation of HL-60 cells over-expressing Ape1/ref-1 was significantly enhanced compared to wild type HL-60 cells. To further understand the mechanism of this effect we performedgel shift experiments in vitro with Ape1/ref-1, retinoic acid receptor alpha (RAR-α), and a retinoic acid response element (RARE) under varying redox conditions andco-transfection experiments in CV-1 cells with Ape1/ref-1 and RAR-α using an RARE linked to a luciferase reporter. Results:gel shift experiments demonstrate a redox dependent binding of RXR-α and RAR-α to their RARE which is mediated by Ape1/ref-1;western blot analysis of transfected CV-1 cells revealed proper expression of each transfected construct including RAR-α, RXR-α and Ape1/ref-1; andexamination of RA-treated CV-1 cells for RARE-linked luciferase expression demonstrated Ape1/ref-1 enhancement of RAR activated transcription of the luciferase reporter. In conclusion, our data supports the contention that Ape1/ref-1 expression may be important for enhancing RA-induced myeloid differentiation and programmed cell death through a redox based mechanism in transcription of target genes.

scholarly journals Terminal differentiation of human promyelocytic leukemic cells in primary culture in response to retinoic acid

Blood ◽  
1981 ◽  
Vol 57 (6) ◽  
pp. 1000-1004 ◽  
Author(s):  
TR Breitman ◽  
SJ Collins ◽  
BR Keene
Keyword(s):  
Retinoic Acid ◽  
Acute Promyelocytic Leukemia ◽  
Leukemia Cell ◽  
Promyelocytic Leukemia ◽  
Leukemic Cells ◽  
Leukemia Cell Line ◽  
Granulocytic Differentiation ◽  
Myelocytic Leukemia ◽  
Functional Maturation ◽  
Promyelocytic Leukemia Cell Line

Abstract The recent finding that retinoic acid induces terminal granulocytic differentiation of the human promyelocytic leukemia cell line, HL-60, prompted an investigation of the sensitivity to this inducer of human myelocytic leukemia cells in primary suspension culture. Of the 21 leukemic specimens, only cells from the two patients with acute promyelocytic leukemia differentiated in response to retinoic acid. After an incubation period of 5--7 days in 1 microM retinoic acid, the cells from these two patients showed extensive morphological and functional maturation. Thus, because it appears that retinoic acid specifically induces granulocytic differentiation of leukemic promyelocytes, this compound may have therapeutic utility in the treatment of acute promyelocytic leukemia.

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