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Exogenous expression of N-cadherin in breast cancer cells induces cell migration, invasion, and metastasis.

Hazan RB, Phillips GR, Qiao RF, Norton L, Aaronson SA - J. Cell Biol. (2000)

Bottom Line: To determine whether N-cadherin promotes invasion and metastasis, we transfected a weakly metastatic and E-cadherin-expressing breast cancer cell line, MCF-7, with N-cadherin and analyzed the effects on cell migration, invasion, and metastasis.These results demonstrate that N-cadherin promotes motility, invasion, and metastasis even in the presence of the normally suppressive E-cadherin.The increase in MMP-9 production by N-cadherin-expressing cells in response to a growth factor may endow them with a greater ability to penetrate matrix protein barriers, while the increase in their adherence to endothelium may improve their ability to enter and exit the vasculature, two properties that may be responsible for metastasis of N-cadherin-expressing cells.

View Article: PubMed Central - PubMed

Affiliation: The Derald H. Ruttenberg Cancer Center, Mount Sinai School of Medicine of New York University, New York, New York 10029, USA. rhazan@smtplink.mssm.edu

ABSTRACT
E- and N-cadherin are calcium-dependent cell adhesion molecules that mediate cell-cell adhesion and also modulate cell migration and tumor invasiveness. The loss of E-cadherin-mediated adhesion has been shown to play an important role in the transition of epithelial tumors from a benign to an invasive state. However, recent evidence indicates that another member of the cadherin family, N-cadherin, is expressed in highly invasive tumor cell lines that lacked E-cadherin expression. These findings have raised the possibility that N-cadherin contributes to the invasive phenotype. To determine whether N-cadherin promotes invasion and metastasis, we transfected a weakly metastatic and E-cadherin-expressing breast cancer cell line, MCF-7, with N-cadherin and analyzed the effects on cell migration, invasion, and metastasis. Transfected cells expressed both E- and N-cadherin and exhibited homotypic cell adhesion from both molecules. In vitro, N-cadherin-expressing cells migrated more efficiently, showed an increased invasion of Matrigel, and adhered more efficiently to monolayers of endothelial cells. All cells produced low levels of the matrix metalloproteinase MMP-9, which was dramatically upregulated by treatment with FGF-2 only in N-cadherin-expressing cells. Migration and invasion of Matrigel were also greatly enhanced by this treatment. When injected into the mammary fat pad of nude mice, N-cadherin-expressing cells, but not control MCF-7 cells, metastasized widely to the liver, pancreas, salivary gland, omentum, lung, lymph nodes, and lumbar spinal muscle. The expression of both E- and N-cadherin was maintained both in the primary tumors and metastatic lesions. These results demonstrate that N-cadherin promotes motility, invasion, and metastasis even in the presence of the normally suppressive E-cadherin. The increase in MMP-9 production by N-cadherin-expressing cells in response to a growth factor may endow them with a greater ability to penetrate matrix protein barriers, while the increase in their adherence to endothelium may improve their ability to enter and exit the vasculature, two properties that may be responsible for metastasis of N-cadherin-expressing cells.

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N-cadherin promotes adhesion of MCF-7 cells to HUVEC cells. (A) Each panel shows an unstained (and therefore not visible) monolayer of HUVEC cells incubated with control Neo-5 (A), N-cad-5 (B), or N-cad-17 (C) cells labeled with fluorescent dye (Fast diO) (see Materials and Methods). D represents N-cadherin immunoblotting of cell extracts from HUVEC (right) compared with Neo-5 (left) and N-cad-5 (middle) control cells.
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Figure 10: N-cadherin promotes adhesion of MCF-7 cells to HUVEC cells. (A) Each panel shows an unstained (and therefore not visible) monolayer of HUVEC cells incubated with control Neo-5 (A), N-cad-5 (B), or N-cad-17 (C) cells labeled with fluorescent dye (Fast diO) (see Materials and Methods). D represents N-cadherin immunoblotting of cell extracts from HUVEC (right) compared with Neo-5 (left) and N-cad-5 (middle) control cells.

Mentions: Additional factors that may critically affect tumor cell metastasis involve interactions between tumor cells and the endothelium. Since endothelial cells are known to express N-cadherin (Salomon et al. 1992), it is possible that the N-cadherin expressed on the surface of tumor cells might promote homophilic interactions with the endothelium. Thus, we tested the ability of control and N-cadherin–expressing MCF-7 cells to adhere to human endothelial monolayers. Whereas N-cad-5 and to a lesser extent, N-cad-17 cells adhered strongly to endothelial cells (Fig. 10B and Fig. C, respectively), control Neo-5 cells exhibited a much weaker adhesion (Fig. 10 A). To control for the relative expression of N-cadherin in these cells, equal amounts of protein extracts from Neo-5 cells, N-cad-5, and HUVEC cells (Fig. 10 D) were immunoblotted with anti–N-cadherin EC1 antibodies. In contrast to control Neo-5 cells, which do not express N-cadherin, HUVEC cells had detectable amounts of N-cadherin, although lesser than that found in N-cad-5 cells. Thus, the ability of N-cadherin–expressing MCF-7 cells to adhere to N-cadherin–expressing endothelial sheets may facilitate their transit through the vasculature and improve their ability to form metastases. In support of these findings, N-cadherin has been shown to mediate the transmigration of melanoma cells through the endothelium (Sandig et al. 1997; Voura et al. 1998).


Exogenous expression of N-cadherin in breast cancer cells induces cell migration, invasion, and metastasis.

Hazan RB, Phillips GR, Qiao RF, Norton L, Aaronson SA - J. Cell Biol. (2000)

N-cadherin promotes adhesion of MCF-7 cells to HUVEC cells. (A) Each panel shows an unstained (and therefore not visible) monolayer of HUVEC cells incubated with control Neo-5 (A), N-cad-5 (B), or N-cad-17 (C) cells labeled with fluorescent dye (Fast diO) (see Materials and Methods). D represents N-cadherin immunoblotting of cell extracts from HUVEC (right) compared with Neo-5 (left) and N-cad-5 (middle) control cells.
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Figure 10: N-cadherin promotes adhesion of MCF-7 cells to HUVEC cells. (A) Each panel shows an unstained (and therefore not visible) monolayer of HUVEC cells incubated with control Neo-5 (A), N-cad-5 (B), or N-cad-17 (C) cells labeled with fluorescent dye (Fast diO) (see Materials and Methods). D represents N-cadherin immunoblotting of cell extracts from HUVEC (right) compared with Neo-5 (left) and N-cad-5 (middle) control cells.
Mentions: Additional factors that may critically affect tumor cell metastasis involve interactions between tumor cells and the endothelium. Since endothelial cells are known to express N-cadherin (Salomon et al. 1992), it is possible that the N-cadherin expressed on the surface of tumor cells might promote homophilic interactions with the endothelium. Thus, we tested the ability of control and N-cadherin–expressing MCF-7 cells to adhere to human endothelial monolayers. Whereas N-cad-5 and to a lesser extent, N-cad-17 cells adhered strongly to endothelial cells (Fig. 10B and Fig. C, respectively), control Neo-5 cells exhibited a much weaker adhesion (Fig. 10 A). To control for the relative expression of N-cadherin in these cells, equal amounts of protein extracts from Neo-5 cells, N-cad-5, and HUVEC cells (Fig. 10 D) were immunoblotted with anti–N-cadherin EC1 antibodies. In contrast to control Neo-5 cells, which do not express N-cadherin, HUVEC cells had detectable amounts of N-cadherin, although lesser than that found in N-cad-5 cells. Thus, the ability of N-cadherin–expressing MCF-7 cells to adhere to N-cadherin–expressing endothelial sheets may facilitate their transit through the vasculature and improve their ability to form metastases. In support of these findings, N-cadherin has been shown to mediate the transmigration of melanoma cells through the endothelium (Sandig et al. 1997; Voura et al. 1998).

Bottom Line: To determine whether N-cadherin promotes invasion and metastasis, we transfected a weakly metastatic and E-cadherin-expressing breast cancer cell line, MCF-7, with N-cadherin and analyzed the effects on cell migration, invasion, and metastasis.These results demonstrate that N-cadherin promotes motility, invasion, and metastasis even in the presence of the normally suppressive E-cadherin.The increase in MMP-9 production by N-cadherin-expressing cells in response to a growth factor may endow them with a greater ability to penetrate matrix protein barriers, while the increase in their adherence to endothelium may improve their ability to enter and exit the vasculature, two properties that may be responsible for metastasis of N-cadherin-expressing cells.

View Article: PubMed Central - PubMed

Affiliation: The Derald H. Ruttenberg Cancer Center, Mount Sinai School of Medicine of New York University, New York, New York 10029, USA. rhazan@smtplink.mssm.edu

ABSTRACT
E- and N-cadherin are calcium-dependent cell adhesion molecules that mediate cell-cell adhesion and also modulate cell migration and tumor invasiveness. The loss of E-cadherin-mediated adhesion has been shown to play an important role in the transition of epithelial tumors from a benign to an invasive state. However, recent evidence indicates that another member of the cadherin family, N-cadherin, is expressed in highly invasive tumor cell lines that lacked E-cadherin expression. These findings have raised the possibility that N-cadherin contributes to the invasive phenotype. To determine whether N-cadherin promotes invasion and metastasis, we transfected a weakly metastatic and E-cadherin-expressing breast cancer cell line, MCF-7, with N-cadherin and analyzed the effects on cell migration, invasion, and metastasis. Transfected cells expressed both E- and N-cadherin and exhibited homotypic cell adhesion from both molecules. In vitro, N-cadherin-expressing cells migrated more efficiently, showed an increased invasion of Matrigel, and adhered more efficiently to monolayers of endothelial cells. All cells produced low levels of the matrix metalloproteinase MMP-9, which was dramatically upregulated by treatment with FGF-2 only in N-cadherin-expressing cells. Migration and invasion of Matrigel were also greatly enhanced by this treatment. When injected into the mammary fat pad of nude mice, N-cadherin-expressing cells, but not control MCF-7 cells, metastasized widely to the liver, pancreas, salivary gland, omentum, lung, lymph nodes, and lumbar spinal muscle. The expression of both E- and N-cadherin was maintained both in the primary tumors and metastatic lesions. These results demonstrate that N-cadherin promotes motility, invasion, and metastasis even in the presence of the normally suppressive E-cadherin. The increase in MMP-9 production by N-cadherin-expressing cells in response to a growth factor may endow them with a greater ability to penetrate matrix protein barriers, while the increase in their adherence to endothelium may improve their ability to enter and exit the vasculature, two properties that may be responsible for metastasis of N-cadherin-expressing cells.

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Related in: MedlinePlus