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Cytotechnology

, Volume 63, Issue 2, pp 101–109 | Cite as

An extra high dose of erythropoietin fails to support the proliferation of erythropoietin dependent cell lines

  • Satoshi Abe
  • Ryuzo Sasaki
  • Seiji MasudaEmail author
JAACT Special Issue

Abstract

Erythropoietin is responsible for the red blood cell formation by stimulating the proliferation and the differentiation of erythroid precursor cells. Erythropoietin triggers the conformational change in its receptor thereby induces the phosphorylation of JAK2. In this study, we show that an extra high dose of erythropoietin, however, fails to activate the erythropoietin receptor, to stimulate the phosphorylation of JAK2 and to support the cell proliferation of Ep-FDC-P2 cell. Moreover, high dose of EPO also inhibited the proliferation of various erythropoietin-dependent cell lines, suggesting that excess amount of EPO could not trigger the conformational change of the receptor. In the presence of an extra high dose of erythropoietin as well as in the absence of erythropoietin, the cells caused the DNA fragmentation, a typical symptom of apoptosis. The impairment of cell growth and the DNA fragmentation at the extremely high concentration of EPO was rescued by the addition of erythropoietin antibody or soluble form of erythropoietin receptor by titrating the excess erythropoietin. These results suggest that two erythropoietin binding sites on erythropoietin receptor dimer should be occupied by a single erythropoietin molecule for the proper conformational change of the receptor and the signal transduction of erythropoietin, instead, when two erythropoietin binding sites on the receptor are shared by two erythropoietin molecules, it fails to evoke the conformational change of erythropoietin receptor adequate for signal transduction.

Keywords

Erythropoietin Erythropoietin receptor JAK2 Apoptosis Antibody Phosphorylation Proliferation Fragmentation 

Abbreviations

EPO

Erythropoietin

EPOR

EPO receptor

sEPOR

Soluble form of EPOR

CFU-E

Colony-forming unit erythroid

IL-3

Interleukin-3

MEM

Minimum essential medium

FCS

Fetal calf serum

MTX

Methotrexate

MTT

3-(4,5-dimethythiazol-2-yl)-2, 5-diphenyl tetrazolium bromide

JAK

Janus family of nonreceptor-type protein tyrosine kinase

STAT

Signal transducers and activators of transcription

Notes

Acknowledgments

This work was supported by Grants-in aid from the Ministry of education, Science and Culture of Japan and from Takeda Science Foundation. We are grateful to Naoko Fujiwara for the fruitful discussion and critical comments.

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  1. 1.Department of Food Science and Technology, Faculty of AgricultureKyoto UniversityKyotoJapan
  2. 2.Graduate School of BiostudiesKyoto UniversityKyotoJapan

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