ABSTRACT
Resident macrophages in bone play important roles in bone remodeling, repair, and hematopoietic stem cell maintenance, yet their role in skeletal metastasis remains under investigated. The purpose of this study was to determine the role of macrophages in prostate cancer skeletal metastasis, using two in vivo mouse models of conditional macrophage depletion. RM-1 syngeneic tumor growth was analyzed in an inducible macrophage (CSF-1 receptor positive cells) ablation model (MAFIA mice). There was a significant reduction in tumor growth in the tibiae of macrophage-ablated mice, compared with control non-ablated mice. Similar results were observed when macrophage ablation was performed using liposome-encapsulated clodronate and human PC-3 prostate cancer cells where tumor-bearing long bones had increased numbers of tumor associated-macrophages. Although tumors were consistently smaller in macrophage-depleted mice, paradoxical results of macrophage depletion on bone were observed. Histomorphometric and micro-CT analyses demonstrated that clodronate-treated mice had increased bone volume, while MAFIA mice had reduced bone volume. These results suggest that the effect of macrophage depletion on tumor growth was independent of its effect on bone responses and that macrophages in bone may be more important to tumor growth than the bone itself. In conclusion, resident macrophages play a pivotal role in prostate cancer growth in bone.
Subject(s)
Bone Marrow Cells/immunology , Bone Neoplasms/immunology , Macrophages/immunology , Prostatic Neoplasms/immunology , Animals , Bone Neoplasms/secondary , Bone Regeneration , Carcinogenesis , Cell Growth Processes , Cell Line, Tumor , Clodronic Acid/metabolism , Humans , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , Prostatic Neoplasms/pathology , Receptor, Macrophage Colony-Stimulating Factor/metabolism , Tumor BurdenABSTRACT
Tumor cells secrete factors that modulate macrophage activation and polarization into M2 type tumor-associated macrophages, which promote tumor growth, progression, and metastasis. The mechanisms that mediate this polarization are not clear. Macrophages are phagocytic cells that participate in the clearance of apoptotic cells, a process known as efferocytosis. Milk fat globule- EGF factor 8 (MFG-E8) is a bridge protein that facilitates efferocytosis and is associated with suppression of proinflammatory responses. This study investigated the hypothesis that MFG-E8-mediated efferocytosis promotes M2 polarization. Tissue and serum exosomes from prostate cancer patients presented higher levels of MFG-E8 compared with controls, a novel finding in human prostate cancer. Coculture of macrophages with apoptotic cancer cells increased efferocytosis, elevated MFG-E8 protein expression levels, and induced macrophage polarization into an alternatively activated M2 phenotype. Administration of antibody against MFG-E8 significantly attenuated the increase in M2 polarization. Inhibition of STAT3 phosphorylation using the inhibitor Stattic decreased efferocytosis and M2 macrophage polarization in vitro, with a correlating increase in SOCS3 protein expression. Moreover, MFG-E8 knockdown tumor cells cultured with wild-type or MFG-E8-deficient macrophages resulted in increased SOCS3 expression with decreased STAT3 activation. This suggests that SOCS3 and phospho-STAT3 act in an inversely dependent manner when stimulated by MFG-E8 and efferocytosis. These results uncover a unique role of efferocytosis via MFG-E8 as a mechanism for macrophage polarization into tumor-promoting M2 cells.
Subject(s)
Antigens, Surface/physiology , Macrophages/immunology , Prostatic Neoplasms/pathology , Animals , Cell Line, Tumor , Flow Cytometry , Male , Mice , Mice, Inbred C57BL , Milk Proteins , Prostatic Neoplasms/immunology , Real-Time Polymerase Chain ReactionABSTRACT
Leuconostoc citreum HO12 and Weissella koreensis HO20 isolated from kimchi were evaluated as starter cultures in the making of whole wheat sourdough bread. After 24h of fermentation at 25 °C, both lactobacilli grew to the final cell numbers of ca. 10(9)cfu/g dough, and both doughs had similar pHs and total titratable acidities. In addition, the fermentation quotient of the dough with Lc. citreum HO12 was slightly lower than that of the dough with W. koreensis HO20 (1.6 versus 2.8). Sourdoughs and bread with 50% sourdough produced with the starter cultures exhibited consistent ability to retard the growth of bread spoilage fungi (Penicillium roqueforti and Aspergillus niger) and rope-forming bacterium (Bacillus subtilis). Sourdough breads underwent a significant reduction in bread firming during storage. It seems that both lactobacilli have the potential to improve the shelf-life of wheat bread. The results indicate that the selected lactobacilli have unique fermentation characteristics and produce sourdough breads with overall satisfactory quality.
