Thec-kit receptor transduces the stem cell factor-triggered growth signal in murine interleukin-3-dependent cell line |
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Authors: | Hideya Ohashi Rie Kameda Mitsuo Nishikawa Mayumi Kawagishi Yun-Cai Liu |
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Institution: | (1) Pharmaceutical Research Laboratory, Kirin Brewery Co., Ltd., 1-2-2, Souja-machi, 371 Maebashi Gunma, Japan |
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Abstract: | The stem cell factor is a glycoprotein hormone which regulates the proliferation and differentiation of primitive hematopoietic cells through its interaction with a tyrosine kinase transmembrane receptor which is encoded by thec-kit proto-oncogene. To examine whether a murinec-kit receptor can be functional in murine interleukin-3 (mlL-3)-dependent hematopoietic cell line, we introduced the murinec-kit cDNA into mlL-3-dependent pro-B cell line Ba/F3. One of the resulting clones, Ba/F3 clone BF-K96, expressed the 140 kDa protein recognized by anti-c-kit monoclonal antibody and the expressedc-kit receptor protein on the cell surface bound to a radiolabeled soluble form of murine stem cell factor (mSCF) with high affinity. BF-K96 clone expressing thec-kit receptor could proliferate in response to mSCF in the absence of mlL-3. The cell clone could also grow in co-culture with mouse 3T3 cells which are endogeneously expressing a membrane-associated type of mSCF on their cell surfaces. These findings demonstrate that thec-kit receptor expressed on mlL-3-dependent hematopoietic cell line Ba/F3 transduce the mSCF-dependent growth signal, indicating that established cell clone will provide a unique cellular system for the study of SCF/c-kit signal transduction mechanism.Abbreviations SCF
stem cell factor
- IL
interleukin
- CSF
colony stimulating factor
- IMDM
Iscove's Modified Dulbecco's Medium
- DME
Dulbecco's Modified Eagle's Medium
- FCS
fetal calf serum
- PCR
polymerase chain reaction
- EDTA
ethylenediaminetetra-acetic acid; sodium dodecyl sulfate |
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