Ohashi H, Kameda R, Nishikawa M, Kawagishi M, Liu Y C
Pharmaceutical Research Laboratory, Kirin Brewery Co., Ltd., Gunma, Japan.
Cytotechnology. 1994;16(1):27-35. doi: 10.1007/BF00761776.
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 the c-kit proto-oncogene. To examine whether a murine c-kit receptor can be functional in murine interleukin-3 (mIL-3)-dependent hematopoietic cell line, we introduced the murine c-kit cDNA into mIL-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 expressed c-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 the c-kit receptor could proliferate in response to mSCF in the absence of mIL-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 the c-kit receptor expressed on mIL-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.
干细胞因子是一种糖蛋白激素,它通过与由原癌基因c-kit编码的酪氨酸激酶跨膜受体相互作用,调节原始造血细胞的增殖和分化。为了检测鼠c-kit受体在依赖鼠白细胞介素-3(mIL-3)的造血细胞系中是否具有功能,我们将鼠c-kit cDNA导入依赖mIL-3的前B细胞系Ba/F3中。其中一个所得克隆,Ba/F3克隆BF-K96,表达了可被抗c-kit单克隆抗体识别的140 kDa蛋白,并且细胞表面表达的c-kit受体蛋白与放射性标记的可溶性鼠干细胞因子(mSCF)以高亲和力结合。表达c-kit受体的BF-K96克隆在没有mIL-3的情况下可响应mSCF而增殖。该细胞克隆也可与在其细胞表面内源性表达膜相关型mSCF的小鼠3T3细胞共培养生长。这些发现表明,在依赖mIL-3的造血细胞系Ba/F3上表达的c-kit受体可转导mSCF依赖的生长信号,这表明所建立的细胞克隆将为研究SCF/c-kit信号转导机制提供一个独特的细胞系统。