Introduction of the Erythropoietin Gene to a Factor-Dependent Murine Myeloid Cell Line View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

1989

AUTHORS

K. Akai , M. Ueda , G. Kawanishi , Y. Miura , T. Suda

ABSTRACT

Erythropoietin (Ep) is a glycoprotein which is required for the proliferation and differentiation of late erythroid progenitor cells. The Ep gene was cloned and large quantities of recombinant Ep is available for the investigation of the action mechanism of Ep. Recently, we found that a small fraction (5%) of an interleukin-3 dependent murine myeloid cell line, FDC-P2 was able to respond to Ep. Moreover, we could establish a subclone EP-FDC-P2, which is dependent on Ep for proliferation and survival, from parental cell FDC-P2. To investigate the role of Ep in cell growth, the Ep gene was introduced into FDC-P2 or EP-FDC-P2 with a retrovirus vector, pZIPNeoSV(X)1. Infected FDC-P2 were selected for G-418 resistance, plated in methylcellulose medium and individual single colonies were cloned. Cell lines (FDC-P2/Ep or EP-FDC- P2/Ep) growing in the absence of exogenous growth factors were established. The Southern blot analysis using the probe of Ep cDNA showed that FDC-P2/EP contained a single copy of the integrated pZIPNeoSV(X)1-Ep provirus. The Northern blot analysis revealed the Ep specific RNA transcript in the cell line. The growth of the FDC-P2/EP cell line was inhibited by a monoclonal antibody against recombinant Ep. However, the amount of the antibody to inhibit the cell growth was much larger than that needed to neutralize Ep activity in the culture medium, suggesting that stimulation of growth occurred not only by external interaction of the secreted Ep with receptors, but also by immediate interaction at cell surface or internal interaction. FDC-P2/EP was not able to induce tumor in nude mice, while transfection of IL-3 gene caused FDC-P2 to become tumorigenic. These results indicate that Ep acts as a growth factor in this system as well as IL-3 and that autocrine stimulation with Ep did not result in tumorigenicity. More... »

PAGES

109-116

Book

TITLE

Experimental Hematology Today—1988

ISBN

978-1-4613-8864-7
978-1-4613-8862-3

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/978-1-4613-8862-3_17

DOI

http://dx.doi.org/10.1007/978-1-4613-8862-3_17

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1038610248


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