Enhancement of J6-1 human leukemic cell proliferation by membrane-bound M-CSF through a cell-cell contact mechanism II. Role of an M-CSF receptor-like membrane protein

We have isolated an M-CSF-like membrane-associated growth factor from human leukemic J6-1 cells that can enhance the growth and colony formation of J6-1 cells in vitro. Indirect evidence suggests that this membrane-associated M-CSF-like growth factor may do so by stimulating a corresponding receptor co-expressed on the adjacent J6-1 cells. The objective of this study is to isolate the putative receptor in J6-1 cells by virtue of its ability to bind and thus "block" the growth of J6-1 cells. Based on this approach, we have isolated from the J6-1 cell membrane an inhibitory activity that can inhibit the clonal growth of J6-1 cells. The activity of this inhibitor can be readily neutralized by either anti-M-CSFR MAb or anti-M-CSFR antiserum, suggesting that it is related to M-CSFR, a product of c-fms proto-oncogene. Judging from Sephadex G-200 gel filtration, the molecular weight (MW) of this putative M-CSFR-like inhibitor was estimated to be approx. 150-180 kDa, comparable with that of M-CSFR. The specificity of M-CSFR-like protein to recognize and block membrane-bound M-CSF also was implicated by its ability to upregulate the steady-state levels of c-fms mRNA in J6-1 cells. Besides its antiproliferative activity in vitro, treatment of J6-1 cells with the putative receptor protein before inoculation effectively blocked the growth and tumor formation in vivo by J6-1 cells in a nude mouse model. These findings suggest that the growth and tumor development by J6-1 leukemic cells may involve a contact-mediated "juxtacrine mechanism". (C) 1998 Elsevier Science Ltd. All rights reserved.