SDF-1alpha regulation in breast cancer cells contacting bone marrow stroma is critical for normal hematopoiesis

Abstract

Breast cancer cells (BCCs) show preference for the bone marrow (BM). An animal model showed 2 populations of BCCs in the BM with regard to their cycling states. An in vitro model of early BC entry into BM showed normal hematopoiesis. Here, we show a critical role for BCC-derived SDF-1alpha in hematopoietic regulation. The studies used a coculture of BM stroma and BCCs (cell lines and stage II BCCs). Northern blots and enzyme-linked immunosorbent assay (ELISA) showed gradual decreases in SDF-1alpha production in BCCs as they contact BM stroma, indicating partial microenvironmental effects caused by stroma on the BCCs. SDF-1 knock-down BCCs and increased exogenous SDF-1alpha prevented contact inhibition between BCCs and BM stroma. Contact inhibition was restored with low SDF-1alpha levels. Long-term culture-initiating assays with CD34(+)/CD38(-)/Lin(-) showed normal hematopoiesis provided that SDF-1alpha levels were reduced in BCCs. Gap junctions (connexin-43 [CX-43]) were formed between BCCs and BM stroma, with concomitant interaction between CD34(+)/CD38(-)/Lin(-) and BM stroma but not with the neighboring BCCs. In summary, SDF-1alpha levels are reduced in BCCs that contact BM stroma. The low levels of SDF-1alpha in BCCs regulate interactions between BM stroma and hematopoietic progenitors, consequently facilitating normal hematopoiesis.