PMID- 15850830
OWN - NLM
STAT- MEDLINE
DCOM- 20050627
LR  - 20111117
IS  - 0301-472X (Print)
IS  - 0301-472X (Linking)
VI  - 33
IP  - 5
DP  - 2005 May
TI  - Hypoxia-inducible factor-1 drives the motility of the erythroid progenitor cell 
      line, UT-7/Epo, via autocrine motility factor.
PG  - 531-41
AB  - OBJECTIVE: It is well known that hypoxic stress strongly enhances erythropoiesis, 
      but the effect of hypoxia on erythroid progenitors has not been examined 
      precisely. In the present study, using the erythropoietin-dependent cell line 
      UT-7/Epo, which has characteristics of erythroid progenitors, we investigated a 
      novel role of hypoxia in erythropoiesis. METHODS: UT-7/Epo and four other 
      hematopoietic and lymphoid cell lines (HL-60, THP-1, Raji, and CEM) were cultured 
      in 20%, 5%, or 1% O2. Morphology was observed under a phase-contrast microscope. 
      Cell motility was evaluated using the Transwell migration assay. An analysis of 
      the protein level of hypoxia-inducible factor-1 (HIF-1) alpha and autocrine 
      motility factor (AMF) was conducted using Western blotting and 
      immunocytochemistry, respectively. Reverse transcription polymerase chain 
      reaction was performed to evaluate the expression of AMF mRNA. Human bone marrow 
      stromal cells were used in cocultures with UT-7/Epo. Apoptosis of UT-7/Epo was 
      examined by immunocytochemistry using an antiactive form of caspase 3 antibody. 
      RESULTS: Among the five cell lines, UT-7/Epo exhibited active pseudopodial 
      extension in hypoxia (1% O2), and cell motility was increased. HL-60, THP-1, 
      Raji, and CEM did not show an increase in cell motility even in 1% O2. In 
      addition, expression of the alpha-subunit of HIF-1 was activated by hypoxia, and 
      expression of the mRNA and protein of AMF induced by HIF-1, increasing cell 
      motility, was promoted. The addition of an HIF-1 inhibitor, cadmium chloride 
      (CdCl2), or alpha-ketoglutarate (2-oxoglutarate) decreased the AMF mRNA 
      expression, and an AMF inhibitor, erythrose 4-phosphate, decreased the cell 
      motility. When UT-7/Epo was cocultured with human bone marrow-derived stromal 
      cells that significantly inhibit the apoptosis of UT-7/Epo, the migration of 
      UT-7/Epo under the stromal cells (pseudoemperipolesis) was increased in hypoxia. 
      CONCLUSION: Under hypoxic conditions, erythroid progenitors may exhibit active 
      migration in the bone marrow and the opportunity for contact with stromal cells 
      increases, inhibiting apoptosis.
FAU - Mikami, Makoto
AU  - Mikami M
AD  - Division of Hematology, Department of Medicine, Kawasaki Medical School, 
      Kurashiki, Japan.
FAU - Sadahira, Yoshito
AU  - Sadahira Y
FAU - Haga, Arayo
AU  - Haga A
FAU - Otsuki, Takemi
AU  - Otsuki T
FAU - Wada, Hideho
AU  - Wada H
FAU - Sugihara, Takashi
AU  - Sugihara T
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Exp Hematol
JT  - Experimental hematology
JID - 0402313
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (HIF1A protein, human)
RN  - 0 (Hypoxia-Inducible Factor 1)
RN  - 0 (Hypoxia-Inducible Factor 1, alpha Subunit)
RN  - 0 (Nuclear Proteins)
RN  - 0 (RNA, Messenger)
RN  - 0 (Transcription Factors)
RN  - EC 5.3.1.9 (Glucose-6-Phosphate Isomerase)
SB  - IM
MH  - Blotting, Western
MH  - Cell Line
MH  - Cell Movement/*physiology
MH  - Coculture Techniques
MH  - DNA-Binding Proteins/*physiology
MH  - Flow Cytometry
MH  - Glucose-6-Phosphate Isomerase/genetics/*physiology
MH  - Humans
MH  - Hypoxia-Inducible Factor 1
MH  - Hypoxia-Inducible Factor 1, alpha Subunit
MH  - Immunohistochemistry
MH  - Nuclear Proteins/*physiology
MH  - RNA, Messenger/genetics
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Transcription Factors/*physiology
EDAT- 2005/04/27 09:00
MHDA- 2005/06/28 09:00
CRDT- 2005/04/27 09:00
PHST- 2004/06/07 00:00 [received]
PHST- 2005/01/26 00:00 [revised]
PHST- 2005/01/27 00:00 [accepted]
PHST- 2005/04/27 09:00 [pubmed]
PHST- 2005/06/28 09:00 [medline]
PHST- 2005/04/27 09:00 [entrez]
AID - S0301-472X(05)00089-5 [pii]
AID - 10.1016/j.exphem.2005.01.013 [doi]
PST - ppublish
SO  - Exp Hematol. 2005 May;33(5):531-41. doi: 10.1016/j.exphem.2005.01.013.