The origin and nature of stromal osteoclast-like multinucleated giant cells in breast carcinoma: implications for tumour osteolysis and macrophage biology

Abstract

(Fry, 1927;Agnantis & Rosen, 1978;Factor et al., 1977;Holland & van Haelst, 1984;Nielsen & Kiaer, 1985;Tavassoli & Norris, 1986) and tumours in other extraskeletal sites (Andreev et al., 1964;Dorney, 1967;Nishiyama et al., 1972; EshunWilson, 1973;Balogh et al., 1985; Berendt et al., 1985). The origin and nature of these cells and their relationship to osteoclasts and macrophage polykaryons is unknown. They are present in the tumour stroma and are generally thought to represent part of the host reponse to the tumour rather than a type of tumour cell. However, their precise function and pathological significance is yet to be determined.In this study, we have examined the ultrastructure and antigenic phenotype of osteoclast-like stromal multinucleate giant cells (OMGCs) in a case of breast carcinoma. We have employed a wide range of monoclonal antibodies of defined specificity in order to clarify the relationship between OMGCs, tumour cells, the osteoclast and other cells of the mononuclear phagocyte system. In addition, we have functionally assessed whether OMGCs behaved like osteoclasts by observing their response to calcitonin and their ability to resorb bone in the presence and absence of known hormonal factors influencing bone resorption. Our findings have implications for macrophage biology and tumour associated osteolysis. Materials and methodsBovine parathyroid hormone (PTH) (2,500 U mg-1) was provided by Dr J. Zanelli (National Institute for Biological Standards, London, UK) and dissolved (230 IU ml-1) in 1 ml 0.001 % acetic acid in distilled water containing 1 mg ml-I of bovine serum albumin (Sigma, UK) (BSA (Welwyn Garden City, UK) and dissolved in alcohol. Tissue culture medium used throughout was Eagles Minimal Essential Medium (Flow, UK), supplemented with benzyl penicillin 100 IU ml-1 (Glaxo, UK) and streptomycin 100 pg ml-1 (Glaxo) (MEM); this was used alone for cell isolation and supplemented with 10% heat inactivated FCS (Gibco, UK) (MEM/FCS) for subsequent incubations.Blocks of cortical bone were obtained at necropsy from the femoral midshaft of patients who had died without evidence of bone disease. These were cleaned of adherent soft tissues then cut longitudinally into bone slices (5 x 3 x 0.3 mm approx) with a low-speed bone saw using a diamond wheel (Buehler Isomet, IL). The bone slices were ultrasonicated for 15min in sterile distilled water, washed in acetone and ethanol then stored dry at room temperature.Anorganic bone slices were prepared by placing bone slices in 5 ml of hot (55°C) 1,2 ethanediamine (Hydrazine) (Eastman, UK). The solution was changed after one hour and the bone slices then kept in hot hydrazine overnight (Termine et al., 1973). The bone slices were washed several times in alcohol then left overnight in alcohol. They were then washed in distilled water, dried and transferred to a sterile petri dish and kept at room temperature until used.Patient details A 45-year-old woman had noted a lump in the upper outer quadrant of her left breast which had recently increased in s...

Bone resorption by macrophage polykaryons of giant cell tumour of tendon sheath

“…In this way, giant cells derived from a giant cell reparative granuloma of the jaw (Flanagan & Chambers, 1988) and giant cells from a giant cell rich malignant fibrous histiocytoma (Flanagan & Chambers, 1989) were considered to be osteoclasts. However, it has recently been shown that tumour-associated macrophage polykaryons derived from a primary breast carcinoma are also able to resorb bone (Athanasou et al, 1989). This result appeared inconsistent with the general belief that no polykaron other than an osteoclast is capable of bone resorption (Chambers & Horton, 1984).…”

“…This pattern of bone resorption is completely different from that of rodent osteoclasts or the osteoclast-like giant cells of a giant cell tumour of bone , both of which produce numerous resorption pits on bone slices. Moreover, although a few large resorption areas were seen, the resorption pits produced by GCTTS and tumour-associated giant cells were generally smaller than those associated with osteoclasts as well as fewer in number (Flanagan & Chambers, 1988;Flanagan & Chambers, 1989;Athanasou et al, 1989). In addition, cells isolated from the GCTTS produced poorly defined areas of surface roughening, a type of bone degradation which has previously been noted to be an uncommon form of osteoclastic bone resorption .…”

Human tumour-associated macrophages are capable of bone resorption

“…Extensive low-grade surface resorption by the numerous mononuclear phagocytes isolated in these studies may be the functional and morphological correlate of this phenomenon. We have previously shown that macrophages and macrophage polykaryons isolated from extraskeletal tumours containing large numbers of these cells are also capable of bone resorption (Athanasou et al, 1989;Athanasou, 1991b); such resorption is similar in type to that described in this study with formation of a few resorption pits and extensive lowgrade surface resorption.…”

“…TAM or AM-containing cell suspensions in alpha minimal essential medium plus 10% foetal calf serum (Gibco) (MEM/FCS) were added to 6mm wells (500cells/well) containing either bone slices or glass coverslips, half of which had been seeded (1,000 cells/well) 24 h earlier with the mouse marrow stromal cell line ST2 (Riken cell bank, Japan) (Athanasou et al, 1989). Four bone slices (half previously seeded with ST2 cells) were studied for each variable, and cell cultures were incubated at 37°C (5% CO2).…”

“…This is largely because direct evidence of bone resorption by these cells has not clearly been demonstrated. However, there is now increasing evidence to show that mononuclear phagocytes derived from inflammatory and neoplastic lesions are capable of bone resorption (Athanasou et al, 1989;Athanasou et al, 1991;Athanasou et al, 1992), and that osteoclast-like cells can form directly from tissue macrophages (Udagawa et al, 1990). …”