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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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Histological view of coculture of mouse calvaria and MDA-231 breast cancer cells embedded in collagen gel. Calvariae were collected from 5-day-old mice, and cocultured for 5 days with MDA-231 cells, embedded in type I collagen gel to retain the position in culture plates. After the culture, the calvariae were fixed, sectioned and stained for HE and TRAP. (A, B) Section of the control calvaria. (C, D) Section of the coculture of calvaria and MDA-231 cells embedded in the gel. Osteoclasts and resorption pits were observed on the surface of the calvaria in contact with the gel containing MDA-231 cells. (E, F) As the positive control, calvariae were cultured for 5 days in the presence of 2 ng ml−1 of IL-1. Osteocalsts were observed inside the calvaria. Cal, calvaria; OC, osteoclasts; Gel, type I collagen gel; MDA, MDA-MD-231.
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fig3: Histological view of coculture of mouse calvaria and MDA-231 breast cancer cells embedded in collagen gel. Calvariae were collected from 5-day-old mice, and cocultured for 5 days with MDA-231 cells, embedded in type I collagen gel to retain the position in culture plates. After the culture, the calvariae were fixed, sectioned and stained for HE and TRAP. (A, B) Section of the control calvaria. (C, D) Section of the coculture of calvaria and MDA-231 cells embedded in the gel. Osteoclasts and resorption pits were observed on the surface of the calvaria in contact with the gel containing MDA-231 cells. (E, F) As the positive control, calvariae were cultured for 5 days in the presence of 2 ng ml−1 of IL-1. Osteocalsts were observed inside the calvaria. Cal, calvaria; OC, osteoclasts; Gel, type I collagen gel; MDA, MDA-MD-231.

Mentions: To examine the mechanism of tumour-induced bone resorption, we developed a new in vitro system using MDA-231 cells and calvaria collected from 5-day-old mice. MDA-231 cells were embedded in type I collagen gel to retain the position and cocultured for 5 days with mouse calvaria. Histological analysis showed that some MDA-231 cells in the collagen gel were in contact with the surface of the calvaria and that TRAP-positive osteoclasts formed resorption pits in the side of the calvaria attached to MDA-231 cells (Figure 3C, DFigure 3


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)

Histological view of coculture of mouse calvaria and MDA-231 breast cancer cells embedded in collagen gel. Calvariae were collected from 5-day-old mice, and cocultured for 5 days with MDA-231 cells, embedded in type I collagen gel to retain the position in culture plates. After the culture, the calvariae were fixed, sectioned and stained for HE and TRAP. (A, B) Section of the control calvaria. (C, D) Section of the coculture of calvaria and MDA-231 cells embedded in the gel. Osteoclasts and resorption pits were observed on the surface of the calvaria in contact with the gel containing MDA-231 cells. (E, F) As the positive control, calvariae were cultured for 5 days in the presence of 2 ng ml−1 of IL-1. Osteocalsts were observed inside the calvaria. Cal, calvaria; OC, osteoclasts; Gel, type I collagen gel; MDA, MDA-MD-231.
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Related In: Results  -  Collection

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fig3: Histological view of coculture of mouse calvaria and MDA-231 breast cancer cells embedded in collagen gel. Calvariae were collected from 5-day-old mice, and cocultured for 5 days with MDA-231 cells, embedded in type I collagen gel to retain the position in culture plates. After the culture, the calvariae were fixed, sectioned and stained for HE and TRAP. (A, B) Section of the control calvaria. (C, D) Section of the coculture of calvaria and MDA-231 cells embedded in the gel. Osteoclasts and resorption pits were observed on the surface of the calvaria in contact with the gel containing MDA-231 cells. (E, F) As the positive control, calvariae were cultured for 5 days in the presence of 2 ng ml−1 of IL-1. Osteocalsts were observed inside the calvaria. Cal, calvaria; OC, osteoclasts; Gel, type I collagen gel; MDA, MDA-MD-231.
Mentions: To examine the mechanism of tumour-induced bone resorption, we developed a new in vitro system using MDA-231 cells and calvaria collected from 5-day-old mice. MDA-231 cells were embedded in type I collagen gel to retain the position and cocultured for 5 days with mouse calvaria. Histological analysis showed that some MDA-231 cells in the collagen gel were in contact with the surface of the calvaria and that TRAP-positive osteoclasts formed resorption pits in the side of the calvaria attached to MDA-231 cells (Figure 3C, DFigure 3

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