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Testes-specific protease 50 promotes cell invasion and metastasis by increasing NF-kappaB-dependent matrix metalloproteinase-9 expression.

Song ZB, Ni JS, Wu P, Bao YL, Liu T, Li M, Fan C, Zhang WJ, Sun LG, Huang YX, Li YX - Cell Death Dis (2015)

Bottom Line: Mechanistic studies revealed that NF-κB signaling pathway was required for TSP50-induced cell migration and metastasis, and further results indicated that TSP50 overexpression enhanced expression and secretion of MMP9, a target gene of NF-κB signaling.In addition, knockdown of MMP9 resulted in inhibition of cell migration and invasion in vitro and lung metastasis in vivo.Furthermore, we found that some breast cancer diagnosis-associated features such as tumor size, tumor grade, estrogen receptors (ER) and progesterone receptors (PR) levels, were correlated well with TSP50/p65 and TSP50/MMP9 expression status.

View Article: PubMed Central - PubMed

Affiliation: 1] National Engineering Laboratory for Druggable Gene and Protein Screening, Northeast Normal University, Changchun 130024, China [2] Research Center of Agriculture and Medicine Gene Engineering of Ministry of Education, Northeast Normal University, Changchun 130024, China.

ABSTRACT
The high mortality in breast cancer is often associated with metastatic progression in patients. Previously we have demonstrated that testes-specific protease 50 (TSP50), an oncogene overexpressed in breast cancer samples, could promote cell proliferation and tumorigenesis. However, whether TSP50 also has a key role in cell invasion and cancer metastasis, and the mechanism underlying the process are still unclear. Here we found that TSP50 overexpression greatly promoted cell migration, invasion, adhesion and formation of the stellate structures in 3D culture system in vitro as well as lung metastasis in vivo. Conversely, TSP50 knockdown caused the opposite changes. Mechanistic studies revealed that NF-κB signaling pathway was required for TSP50-induced cell migration and metastasis, and further results indicated that TSP50 overexpression enhanced expression and secretion of MMP9, a target gene of NF-κB signaling. In addition, knockdown of MMP9 resulted in inhibition of cell migration and invasion in vitro and lung metastasis in vivo. Most importantly, immunohistochemical staining of human breast cancer samples strongly showed that the coexpression of TSP50 and p65 as well as TSP50 and MMP9 were correlated with increased metastasis and poor survival. Furthermore, we found that some breast cancer diagnosis-associated features such as tumor size, tumor grade, estrogen receptors (ER) and progesterone receptors (PR) levels, were correlated well with TSP50/p65 and TSP50/MMP9 expression status. Taken together, this work identified the TSP50 activation of MMP9 as a novel signaling mechanism underlying human breast cancer invasion and metastasis.

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NF-κB activation is required in TSP50-induced cell migration and invasion. (a) Top panel: Overexpression of IκB-SR. Bottom panel: Overexpression of IκB-SR suppressed TSP50-induced p65 nuclear translocation by immunofluorescence assay. (b) Overexpression of IκB-SR suppressed TSP50-induced cell migration. Scale bar, 100 μm. (c) Overexpression of IκB-SR suppressed TSP50-induced cell invasion. Scale bar, 50 μm. (d) Overexpression of IκB-SR suppressed TSP50-induced cell adhesion. Scale bar, 150 μm. (e) Effects of I κB-SR expression on the morphology of TSP50-expressing cells. The arrow points out the filopodium. Scale bar, 50 μm
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fig4: NF-κB activation is required in TSP50-induced cell migration and invasion. (a) Top panel: Overexpression of IκB-SR. Bottom panel: Overexpression of IκB-SR suppressed TSP50-induced p65 nuclear translocation by immunofluorescence assay. (b) Overexpression of IκB-SR suppressed TSP50-induced cell migration. Scale bar, 100 μm. (c) Overexpression of IκB-SR suppressed TSP50-induced cell invasion. Scale bar, 50 μm. (d) Overexpression of IκB-SR suppressed TSP50-induced cell adhesion. Scale bar, 150 μm. (e) Effects of I κB-SR expression on the morphology of TSP50-expressing cells. The arrow points out the filopodium. Scale bar, 50 μm

Mentions: Previously, we have demonstrated that TSP50 promoted cell proliferation and tumor formation through activation of NF-κB signaling pathway.22 To determine whether NF-κB activation was also necessary for TSP50-induced cell migration and invasion, a dominant-negative IκB mutant (IκB-SR) was transfected into control and TSP50-expressing CHO cells to block NF-κB activation as described previously,22 and the expression of IκB-SR and its effects on TSP50-induced p65 nuclear translocation was further determined (Figure 4a). As expect, cell migration of TSP50-expressing cells was reduced by IκB-SR expression (Figure 4b). In addition, IκB-SR expression greatly inhibited TSP50-induced cell invasion by transwell chamber assay (Figure 4c), suggesting that TSP50 required NF-κB signal to enhance cell mobility. Likewise, TSP50-induced cell adhesion and the length of cell stellate structures in 3D medium were significantly reduced (Figures 4d and e). Taken together, these results suggest that NF-κB activation is required in TSP50-induced cell migration and invasion.


Testes-specific protease 50 promotes cell invasion and metastasis by increasing NF-kappaB-dependent matrix metalloproteinase-9 expression.

Song ZB, Ni JS, Wu P, Bao YL, Liu T, Li M, Fan C, Zhang WJ, Sun LG, Huang YX, Li YX - Cell Death Dis (2015)

NF-κB activation is required in TSP50-induced cell migration and invasion. (a) Top panel: Overexpression of IκB-SR. Bottom panel: Overexpression of IκB-SR suppressed TSP50-induced p65 nuclear translocation by immunofluorescence assay. (b) Overexpression of IκB-SR suppressed TSP50-induced cell migration. Scale bar, 100 μm. (c) Overexpression of IκB-SR suppressed TSP50-induced cell invasion. Scale bar, 50 μm. (d) Overexpression of IκB-SR suppressed TSP50-induced cell adhesion. Scale bar, 150 μm. (e) Effects of I κB-SR expression on the morphology of TSP50-expressing cells. The arrow points out the filopodium. Scale bar, 50 μm
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4385939&req=5

fig4: NF-κB activation is required in TSP50-induced cell migration and invasion. (a) Top panel: Overexpression of IκB-SR. Bottom panel: Overexpression of IκB-SR suppressed TSP50-induced p65 nuclear translocation by immunofluorescence assay. (b) Overexpression of IκB-SR suppressed TSP50-induced cell migration. Scale bar, 100 μm. (c) Overexpression of IκB-SR suppressed TSP50-induced cell invasion. Scale bar, 50 μm. (d) Overexpression of IκB-SR suppressed TSP50-induced cell adhesion. Scale bar, 150 μm. (e) Effects of I κB-SR expression on the morphology of TSP50-expressing cells. The arrow points out the filopodium. Scale bar, 50 μm
Mentions: Previously, we have demonstrated that TSP50 promoted cell proliferation and tumor formation through activation of NF-κB signaling pathway.22 To determine whether NF-κB activation was also necessary for TSP50-induced cell migration and invasion, a dominant-negative IκB mutant (IκB-SR) was transfected into control and TSP50-expressing CHO cells to block NF-κB activation as described previously,22 and the expression of IκB-SR and its effects on TSP50-induced p65 nuclear translocation was further determined (Figure 4a). As expect, cell migration of TSP50-expressing cells was reduced by IκB-SR expression (Figure 4b). In addition, IκB-SR expression greatly inhibited TSP50-induced cell invasion by transwell chamber assay (Figure 4c), suggesting that TSP50 required NF-κB signal to enhance cell mobility. Likewise, TSP50-induced cell adhesion and the length of cell stellate structures in 3D medium were significantly reduced (Figures 4d and e). Taken together, these results suggest that NF-κB activation is required in TSP50-induced cell migration and invasion.

Bottom Line: Mechanistic studies revealed that NF-κB signaling pathway was required for TSP50-induced cell migration and metastasis, and further results indicated that TSP50 overexpression enhanced expression and secretion of MMP9, a target gene of NF-κB signaling.In addition, knockdown of MMP9 resulted in inhibition of cell migration and invasion in vitro and lung metastasis in vivo.Furthermore, we found that some breast cancer diagnosis-associated features such as tumor size, tumor grade, estrogen receptors (ER) and progesterone receptors (PR) levels, were correlated well with TSP50/p65 and TSP50/MMP9 expression status.

View Article: PubMed Central - PubMed

Affiliation: 1] National Engineering Laboratory for Druggable Gene and Protein Screening, Northeast Normal University, Changchun 130024, China [2] Research Center of Agriculture and Medicine Gene Engineering of Ministry of Education, Northeast Normal University, Changchun 130024, China.

ABSTRACT
The high mortality in breast cancer is often associated with metastatic progression in patients. Previously we have demonstrated that testes-specific protease 50 (TSP50), an oncogene overexpressed in breast cancer samples, could promote cell proliferation and tumorigenesis. However, whether TSP50 also has a key role in cell invasion and cancer metastasis, and the mechanism underlying the process are still unclear. Here we found that TSP50 overexpression greatly promoted cell migration, invasion, adhesion and formation of the stellate structures in 3D culture system in vitro as well as lung metastasis in vivo. Conversely, TSP50 knockdown caused the opposite changes. Mechanistic studies revealed that NF-κB signaling pathway was required for TSP50-induced cell migration and metastasis, and further results indicated that TSP50 overexpression enhanced expression and secretion of MMP9, a target gene of NF-κB signaling. In addition, knockdown of MMP9 resulted in inhibition of cell migration and invasion in vitro and lung metastasis in vivo. Most importantly, immunohistochemical staining of human breast cancer samples strongly showed that the coexpression of TSP50 and p65 as well as TSP50 and MMP9 were correlated with increased metastasis and poor survival. Furthermore, we found that some breast cancer diagnosis-associated features such as tumor size, tumor grade, estrogen receptors (ER) and progesterone receptors (PR) levels, were correlated well with TSP50/p65 and TSP50/MMP9 expression status. Taken together, this work identified the TSP50 activation of MMP9 as a novel signaling mechanism underlying human breast cancer invasion and metastasis.

Show MeSH
Related in: MedlinePlus