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Role of RANKL-induced osteoclast formation and MMP-dependent matrix degradation in bone destruction by breast cancer metastasis.

Ohshiba T, Miyaura C, Inada M, Ito A - Br. J. Cancer (2003)

Bottom Line: The separation of MDA-231 from the calvaria using filter insert showed decreased bone resorption, suggesting that cell-to-cell interaction is essential for cancer-induced bone resorption.Adding MDA-231 cells to bone marrow cultures markedly induced osteoclast formation, and the expression of RANKL in osteoblasts was enhanced by contact with the cell surface of MDA-231 cells.These results indicate that RANKL-induced osteoclast formation and MMP-dependent matrix degradation are associated with osteolysis because of bone metastasis of breast cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, School of Pharmacy, Tokyo University of Pharmacy and Life Science, Tokyo, Japan.

ABSTRACT
Bone metastasis of breast cancer induces severe osteolysis with increased bone resorption. Osteoclast differentiation regulated by the receptor activator of NF-kappaB ligand (RANKL) in osteoblasts and matrix degradation induced by matrix metalloproteinases (MMPs) are thought to be involved in the process of bone resorption. When nude mice were inoculated with human breast cancer cells, MDA-MB-231(MDA-231), numerous osteoclasts resorbed bone and the degradation of the bone matrix markedly progressed in the femur and tibia with metastasis of the MDA-231 tumour. The expression of RANKL, MMP-13 and membrane-type 1-MMP mRNA was markedly elevated in bone with metastasis. When MDA-231 cells were cocultured with mouse calvaria, MDA-231 markedly induced bone resorption measured by calcium release from the calvaria, and the expression of RANKL, MMP-2 and MMP-13 was elevated in the calvaria after the coculture. The separation of MDA-231 from the calvaria using filter insert showed decreased bone resorption, suggesting that cell-to-cell interaction is essential for cancer-induced bone resorption. Adding MDA-231 cells to bone marrow cultures markedly induced osteoclast formation, and the expression of RANKL in osteoblasts was enhanced by contact with the cell surface of MDA-231 cells. These results indicate that RANKL-induced osteoclast formation and MMP-dependent matrix degradation are associated with osteolysis because of bone metastasis of breast cancer.

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Osteoclast formation induced by MDA-231 cells in cocultures of mouse bone marrow cells and osteoblasts. Mouse bone marrow cells (2 × 106) and osteoblasts (1 × 104) were cocultured for 8 days with MDA-231 cells (1 × 104) in the presence or absence of 100 ng ml−1 of OPG. In a separate coculture, MDA-231 cells were separated from bone marrow cells and osteoblasts using a cell culture insert. (A) Representative field of TRAP staining. (B) The number of TRAP-positive multinucleated cells (MNCs) containing three or more nuclei was counted. Data are expressed as the means±s.e.m. of three wells. Significantly different from the control (*P<0.001).
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fig6: Osteoclast formation induced by MDA-231 cells in cocultures of mouse bone marrow cells and osteoblasts. Mouse bone marrow cells (2 × 106) and osteoblasts (1 × 104) were cocultured for 8 days with MDA-231 cells (1 × 104) in the presence or absence of 100 ng ml−1 of OPG. In a separate coculture, MDA-231 cells were separated from bone marrow cells and osteoblasts using a cell culture insert. (A) Representative field of TRAP staining. (B) The number of TRAP-positive multinucleated cells (MNCs) containing three or more nuclei was counted. Data are expressed as the means±s.e.m. of three wells. Significantly different from the control (*P<0.001).

Mentions: In place of osteolysis because of bone metastasis, numerous osteoclasts appear on the surface of bone in vivo, as shown in Figure 1. To test whether osteoclasts are formed by the presence of tumour cells in vitro, we added MDA-231 cells to the coculture of mouse bone marrow cells and osteoblasts. Usually, the addition of bone-resorbing factors is essential for osteoclast formation in the coculture system. However, as shown in Figure 6Figure 6


Role of RANKL-induced osteoclast formation and MMP-dependent matrix degradation in bone destruction by breast cancer metastasis.

Ohshiba T, Miyaura C, Inada M, Ito A - Br. J. Cancer (2003)

Osteoclast formation induced by MDA-231 cells in cocultures of mouse bone marrow cells and osteoblasts. Mouse bone marrow cells (2 × 106) and osteoblasts (1 × 104) were cocultured for 8 days with MDA-231 cells (1 × 104) in the presence or absence of 100 ng ml−1 of OPG. In a separate coculture, MDA-231 cells were separated from bone marrow cells and osteoblasts using a cell culture insert. (A) Representative field of TRAP staining. (B) The number of TRAP-positive multinucleated cells (MNCs) containing three or more nuclei was counted. Data are expressed as the means±s.e.m. of three wells. Significantly different from the control (*P<0.001).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2747560&req=5

fig6: Osteoclast formation induced by MDA-231 cells in cocultures of mouse bone marrow cells and osteoblasts. Mouse bone marrow cells (2 × 106) and osteoblasts (1 × 104) were cocultured for 8 days with MDA-231 cells (1 × 104) in the presence or absence of 100 ng ml−1 of OPG. In a separate coculture, MDA-231 cells were separated from bone marrow cells and osteoblasts using a cell culture insert. (A) Representative field of TRAP staining. (B) The number of TRAP-positive multinucleated cells (MNCs) containing three or more nuclei was counted. Data are expressed as the means±s.e.m. of three wells. Significantly different from the control (*P<0.001).
Mentions: In place of osteolysis because of bone metastasis, numerous osteoclasts appear on the surface of bone in vivo, as shown in Figure 1. To test whether osteoclasts are formed by the presence of tumour cells in vitro, we added MDA-231 cells to the coculture of mouse bone marrow cells and osteoblasts. Usually, the addition of bone-resorbing factors is essential for osteoclast formation in the coculture system. However, as shown in Figure 6Figure 6

Bottom Line: The separation of MDA-231 from the calvaria using filter insert showed decreased bone resorption, suggesting that cell-to-cell interaction is essential for cancer-induced bone resorption.Adding MDA-231 cells to bone marrow cultures markedly induced osteoclast formation, and the expression of RANKL in osteoblasts was enhanced by contact with the cell surface of MDA-231 cells.These results indicate that RANKL-induced osteoclast formation and MMP-dependent matrix degradation are associated with osteolysis because of bone metastasis of breast cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, School of Pharmacy, Tokyo University of Pharmacy and Life Science, Tokyo, Japan.

ABSTRACT
Bone metastasis of breast cancer induces severe osteolysis with increased bone resorption. Osteoclast differentiation regulated by the receptor activator of NF-kappaB ligand (RANKL) in osteoblasts and matrix degradation induced by matrix metalloproteinases (MMPs) are thought to be involved in the process of bone resorption. When nude mice were inoculated with human breast cancer cells, MDA-MB-231(MDA-231), numerous osteoclasts resorbed bone and the degradation of the bone matrix markedly progressed in the femur and tibia with metastasis of the MDA-231 tumour. The expression of RANKL, MMP-13 and membrane-type 1-MMP mRNA was markedly elevated in bone with metastasis. When MDA-231 cells were cocultured with mouse calvaria, MDA-231 markedly induced bone resorption measured by calcium release from the calvaria, and the expression of RANKL, MMP-2 and MMP-13 was elevated in the calvaria after the coculture. The separation of MDA-231 from the calvaria using filter insert showed decreased bone resorption, suggesting that cell-to-cell interaction is essential for cancer-induced bone resorption. Adding MDA-231 cells to bone marrow cultures markedly induced osteoclast formation, and the expression of RANKL in osteoblasts was enhanced by contact with the cell surface of MDA-231 cells. These results indicate that RANKL-induced osteoclast formation and MMP-dependent matrix degradation are associated with osteolysis because of bone metastasis of breast cancer.

Show MeSH
Related in: MedlinePlus