Accumulation of hypoxia-inducible factor-1α protein and its role in the differentiation of myeloid leukemic cells induced by all-trans retinoic acid

Jing Zhang, Li Ping Song, Ying Huang, Qian Zhao, Ke Wen Zhao, Guo Qiang Chen

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Background: The clinical activities of all-trans retinoic acid in the treatment of acute promyelocytic leukemia, a unique subtype of acute myeloid leukemia, have triggered extensive studies aimed at defining the mechanisms by which this compound induces differentiation of leukemic cells. Recent studies show that hypoxia-inducible factor-1α (HIF-1α) contributes to the differentiation of acute myeloid leukemia cells via transcriptional activity-independent mechanisms. We investigated whether all-trans retinoic acid affects HIF-1α protein and whether this has a role in all-trans retinoic acid-induced differentiation. Design and Methods: The acute myeloid leukemia cell lines NB4 and U937 were treated with all-trans retinoic acid, and HIF-1α/HIF-1β mRNA and proteins were measured respectively by real-time quantitative reverse transcriptase polymerase chain reaction and western blotting. To investigate the role of HIF-1α in all-trans retinoic acid-induced differentiation, NB4 cells, U937 cells, U937 cells in which HIF-1α was induced by the withdrawal of tetracycline and U937 cells with stable expression of specific short hairpin RNA against HIF-1α, Runx1, C/EBPα and PU.1, were treated with all-trans retinoic acid and/or the hypoxia-mimetic agent cobalt chloride (CoCl2). Cellular differentiation was evaluated by morphological criteria and myeloid differentiation antigens. Results: all-trans retinoic acid rapidly increased endogenous and inducible expressed or CoCl2-stabilized HIF-1α protein in leukemic cells under normoxia. Importantly, suppression of HIF-1α expression by specific short hairpin RNA partially but significantly inhibited all-trans retinoic acid-induced differentiation of the U937 cell line. Reciprocally, the differentiation induced by all-trans retinoic acid was significantly enhanced by conditional HIF-1α induction and HIF-1α-stabilizing CoCl2 treatment. Furthermore, knock-down of PU.1, Runx1 and C/EBPα, three transcriptional factors crucial for normal hematopoiesis, greatly inhibited the differentiation cooperation of all-trans retinoic acid and HIF-1α induction. Conclusions: This work provides the first demonstration that HIF-1α, a protein rapidly responsive to all-trans retinoic acid, plays a role in all-trans retinoic acid-induced differentiation of leukemic cells. These observations shed new light on the molecular mechanisms underlying all-trans retinoic acid-induced differentiation of acute myeloid leukemia cells.

Original languageEnglish (US)
Pages (from-to)1480-1487
Number of pages8
JournalHaematologica
Volume93
Issue number10
DOIs
StatePublished - Oct 1 2008

Fingerprint

Hypoxia-Inducible Factor 1
Myeloid Cells
Tretinoin
Proteins
U937 Cells
Acute Myeloid Leukemia
Cell Differentiation
Small Interfering RNA
Cell Line
Acute Promyelocytic Leukemia
Differentiation Antigens
Hematopoiesis
Tetracycline
Reverse Transcriptase Polymerase Chain Reaction

Keywords

  • all-trans retinoid acid
  • Differentiation
  • Hypoxia inducible factor-1α
  • Leukemia

ASJC Scopus subject areas

  • Hematology

Cite this

Accumulation of hypoxia-inducible factor-1α protein and its role in the differentiation of myeloid leukemic cells induced by all-trans retinoic acid. / Zhang, Jing; Song, Li Ping; Huang, Ying; Zhao, Qian; Zhao, Ke Wen; Chen, Guo Qiang.

In: Haematologica, Vol. 93, No. 10, 01.10.2008, p. 1480-1487.

Research output: Contribution to journalArticle

Zhang, Jing ; Song, Li Ping ; Huang, Ying ; Zhao, Qian ; Zhao, Ke Wen ; Chen, Guo Qiang. / Accumulation of hypoxia-inducible factor-1α protein and its role in the differentiation of myeloid leukemic cells induced by all-trans retinoic acid. In: Haematologica. 2008 ; Vol. 93, No. 10. pp. 1480-1487.
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abstract = "Background: The clinical activities of all-trans retinoic acid in the treatment of acute promyelocytic leukemia, a unique subtype of acute myeloid leukemia, have triggered extensive studies aimed at defining the mechanisms by which this compound induces differentiation of leukemic cells. Recent studies show that hypoxia-inducible factor-1α (HIF-1α) contributes to the differentiation of acute myeloid leukemia cells via transcriptional activity-independent mechanisms. We investigated whether all-trans retinoic acid affects HIF-1α protein and whether this has a role in all-trans retinoic acid-induced differentiation. Design and Methods: The acute myeloid leukemia cell lines NB4 and U937 were treated with all-trans retinoic acid, and HIF-1α/HIF-1β mRNA and proteins were measured respectively by real-time quantitative reverse transcriptase polymerase chain reaction and western blotting. To investigate the role of HIF-1α in all-trans retinoic acid-induced differentiation, NB4 cells, U937 cells, U937 cells in which HIF-1α was induced by the withdrawal of tetracycline and U937 cells with stable expression of specific short hairpin RNA against HIF-1α, Runx1, C/EBPα and PU.1, were treated with all-trans retinoic acid and/or the hypoxia-mimetic agent cobalt chloride (CoCl2). Cellular differentiation was evaluated by morphological criteria and myeloid differentiation antigens. Results: all-trans retinoic acid rapidly increased endogenous and inducible expressed or CoCl2-stabilized HIF-1α protein in leukemic cells under normoxia. Importantly, suppression of HIF-1α expression by specific short hairpin RNA partially but significantly inhibited all-trans retinoic acid-induced differentiation of the U937 cell line. Reciprocally, the differentiation induced by all-trans retinoic acid was significantly enhanced by conditional HIF-1α induction and HIF-1α-stabilizing CoCl2 treatment. Furthermore, knock-down of PU.1, Runx1 and C/EBPα, three transcriptional factors crucial for normal hematopoiesis, greatly inhibited the differentiation cooperation of all-trans retinoic acid and HIF-1α induction. Conclusions: This work provides the first demonstration that HIF-1α, a protein rapidly responsive to all-trans retinoic acid, plays a role in all-trans retinoic acid-induced differentiation of leukemic cells. These observations shed new light on the molecular mechanisms underlying all-trans retinoic acid-induced differentiation of acute myeloid leukemia cells.",
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AU - Zhang, Jing

AU - Song, Li Ping

AU - Huang, Ying

AU - Zhao, Qian

AU - Zhao, Ke Wen

AU - Chen, Guo Qiang

PY - 2008/10/1

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N2 - Background: The clinical activities of all-trans retinoic acid in the treatment of acute promyelocytic leukemia, a unique subtype of acute myeloid leukemia, have triggered extensive studies aimed at defining the mechanisms by which this compound induces differentiation of leukemic cells. Recent studies show that hypoxia-inducible factor-1α (HIF-1α) contributes to the differentiation of acute myeloid leukemia cells via transcriptional activity-independent mechanisms. We investigated whether all-trans retinoic acid affects HIF-1α protein and whether this has a role in all-trans retinoic acid-induced differentiation. Design and Methods: The acute myeloid leukemia cell lines NB4 and U937 were treated with all-trans retinoic acid, and HIF-1α/HIF-1β mRNA and proteins were measured respectively by real-time quantitative reverse transcriptase polymerase chain reaction and western blotting. To investigate the role of HIF-1α in all-trans retinoic acid-induced differentiation, NB4 cells, U937 cells, U937 cells in which HIF-1α was induced by the withdrawal of tetracycline and U937 cells with stable expression of specific short hairpin RNA against HIF-1α, Runx1, C/EBPα and PU.1, were treated with all-trans retinoic acid and/or the hypoxia-mimetic agent cobalt chloride (CoCl2). Cellular differentiation was evaluated by morphological criteria and myeloid differentiation antigens. Results: all-trans retinoic acid rapidly increased endogenous and inducible expressed or CoCl2-stabilized HIF-1α protein in leukemic cells under normoxia. Importantly, suppression of HIF-1α expression by specific short hairpin RNA partially but significantly inhibited all-trans retinoic acid-induced differentiation of the U937 cell line. Reciprocally, the differentiation induced by all-trans retinoic acid was significantly enhanced by conditional HIF-1α induction and HIF-1α-stabilizing CoCl2 treatment. Furthermore, knock-down of PU.1, Runx1 and C/EBPα, three transcriptional factors crucial for normal hematopoiesis, greatly inhibited the differentiation cooperation of all-trans retinoic acid and HIF-1α induction. Conclusions: This work provides the first demonstration that HIF-1α, a protein rapidly responsive to all-trans retinoic acid, plays a role in all-trans retinoic acid-induced differentiation of leukemic cells. These observations shed new light on the molecular mechanisms underlying all-trans retinoic acid-induced differentiation of acute myeloid leukemia cells.

AB - Background: The clinical activities of all-trans retinoic acid in the treatment of acute promyelocytic leukemia, a unique subtype of acute myeloid leukemia, have triggered extensive studies aimed at defining the mechanisms by which this compound induces differentiation of leukemic cells. Recent studies show that hypoxia-inducible factor-1α (HIF-1α) contributes to the differentiation of acute myeloid leukemia cells via transcriptional activity-independent mechanisms. We investigated whether all-trans retinoic acid affects HIF-1α protein and whether this has a role in all-trans retinoic acid-induced differentiation. Design and Methods: The acute myeloid leukemia cell lines NB4 and U937 were treated with all-trans retinoic acid, and HIF-1α/HIF-1β mRNA and proteins were measured respectively by real-time quantitative reverse transcriptase polymerase chain reaction and western blotting. To investigate the role of HIF-1α in all-trans retinoic acid-induced differentiation, NB4 cells, U937 cells, U937 cells in which HIF-1α was induced by the withdrawal of tetracycline and U937 cells with stable expression of specific short hairpin RNA against HIF-1α, Runx1, C/EBPα and PU.1, were treated with all-trans retinoic acid and/or the hypoxia-mimetic agent cobalt chloride (CoCl2). Cellular differentiation was evaluated by morphological criteria and myeloid differentiation antigens. Results: all-trans retinoic acid rapidly increased endogenous and inducible expressed or CoCl2-stabilized HIF-1α protein in leukemic cells under normoxia. Importantly, suppression of HIF-1α expression by specific short hairpin RNA partially but significantly inhibited all-trans retinoic acid-induced differentiation of the U937 cell line. Reciprocally, the differentiation induced by all-trans retinoic acid was significantly enhanced by conditional HIF-1α induction and HIF-1α-stabilizing CoCl2 treatment. Furthermore, knock-down of PU.1, Runx1 and C/EBPα, three transcriptional factors crucial for normal hematopoiesis, greatly inhibited the differentiation cooperation of all-trans retinoic acid and HIF-1α induction. Conclusions: This work provides the first demonstration that HIF-1α, a protein rapidly responsive to all-trans retinoic acid, plays a role in all-trans retinoic acid-induced differentiation of leukemic cells. These observations shed new light on the molecular mechanisms underlying all-trans retinoic acid-induced differentiation of acute myeloid leukemia cells.

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KW - Differentiation

KW - Hypoxia inducible factor-1α

KW - Leukemia

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