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Binding of TGF-beta1 latency-associated peptide (LAP) to alpha(v)beta6 integrin modulates behaviour of squamous carcinoma cells.

Thomas GJ, Hart IR, Speight PM, Marshall JF - Br. J. Cancer (2002)

Bottom Line: Since TGF-beta1 is present in squamous carcinomas, it is possible that latency associated peptide may modulate malignant keratinocyte behaviour independently from the classical TGF-beta signalling pathways through its interaction with integrins.We show here that when latency associated peptide is immobilised onto a surface, it acts as an alpha(v)beta6-specific ligand for oral squamous carcinoma cells promoting adhesion and haptotactic migration in addition to alpha(v)beta6-dependent increase in pro-MMP-9 expression.In contrast, even very low concentrations of soluble latency associated peptide (0.1 microg ml(-1)) inhibited alpha(v)beta6-dependent adhesion, migration and invasion.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Pathology, Eastman Dental Institute, University College London, UK.

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Binding of αvβ6 to LAP induces upregulation of MMP-9. Zymography for MMP-9. Cells were grown for 24 h in additive-free medium before supernatant sampling and cell counting. Samples containing equal volume per cell number were run on each gel with MMP-9 control. The intensity of the bands was measured by densitometric analysis and comparisons made within each gel to determine relative changes in MMP activity. (A) Zymogram showing upregulation of MMP-9 expression by VB6 cells when plated on fibronectin (FN) and LAP relative to BSA-coated plastic. Control is pro-MMP-9. (B) Zymogram showing the inhibition of MMP-9 expression by VB6 cells plated onto LAP following blockade with anti-αvβ6 antibody 10D5 relative to an irrelevant control antibody (Ctl; W632 (anti-MHC class 1)). (C) Densitometric analysis of zymograms from multiple separate experiments (n=4) showing MMP-9 expression by VB6 cells on BSA, LAP following incubation with an irrelevant control antibody (Ctl; W632 (anti-MHC class 1)) and LAP following blockade with anti-αvβ6 antibody (10D5). Results are expressed relative to MMP-9 expression by VB6 cells on BSA-coated plastic (=100). Error bars represent standard deviation. These data suggest that αvβ6-LAP interaction modulates MMP-9 expression in VB6 cells.
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fig4: Binding of αvβ6 to LAP induces upregulation of MMP-9. Zymography for MMP-9. Cells were grown for 24 h in additive-free medium before supernatant sampling and cell counting. Samples containing equal volume per cell number were run on each gel with MMP-9 control. The intensity of the bands was measured by densitometric analysis and comparisons made within each gel to determine relative changes in MMP activity. (A) Zymogram showing upregulation of MMP-9 expression by VB6 cells when plated on fibronectin (FN) and LAP relative to BSA-coated plastic. Control is pro-MMP-9. (B) Zymogram showing the inhibition of MMP-9 expression by VB6 cells plated onto LAP following blockade with anti-αvβ6 antibody 10D5 relative to an irrelevant control antibody (Ctl; W632 (anti-MHC class 1)). (C) Densitometric analysis of zymograms from multiple separate experiments (n=4) showing MMP-9 expression by VB6 cells on BSA, LAP following incubation with an irrelevant control antibody (Ctl; W632 (anti-MHC class 1)) and LAP following blockade with anti-αvβ6 antibody (10D5). Results are expressed relative to MMP-9 expression by VB6 cells on BSA-coated plastic (=100). Error bars represent standard deviation. These data suggest that αvβ6-LAP interaction modulates MMP-9 expression in VB6 cells.

Mentions: Expression of MMP-9 on BSA-, fibronectin-, and LAP-coated tissue culture plastic was examined by zymography. Zymography on supernatant samples from cells grown in serum-free medium showed that the cells produced predominantly MMP-9 and that this enzyme was mainly present in proenzyme form. Similar to the effect observed on fibronectin (Thomas et al, 2001c), VB6 cells showed significant upregulation of MMP-9 when plated on LAP (P=<0.001; Figure 4A,CFigure 4


Binding of TGF-beta1 latency-associated peptide (LAP) to alpha(v)beta6 integrin modulates behaviour of squamous carcinoma cells.

Thomas GJ, Hart IR, Speight PM, Marshall JF - Br. J. Cancer (2002)

Binding of αvβ6 to LAP induces upregulation of MMP-9. Zymography for MMP-9. Cells were grown for 24 h in additive-free medium before supernatant sampling and cell counting. Samples containing equal volume per cell number were run on each gel with MMP-9 control. The intensity of the bands was measured by densitometric analysis and comparisons made within each gel to determine relative changes in MMP activity. (A) Zymogram showing upregulation of MMP-9 expression by VB6 cells when plated on fibronectin (FN) and LAP relative to BSA-coated plastic. Control is pro-MMP-9. (B) Zymogram showing the inhibition of MMP-9 expression by VB6 cells plated onto LAP following blockade with anti-αvβ6 antibody 10D5 relative to an irrelevant control antibody (Ctl; W632 (anti-MHC class 1)). (C) Densitometric analysis of zymograms from multiple separate experiments (n=4) showing MMP-9 expression by VB6 cells on BSA, LAP following incubation with an irrelevant control antibody (Ctl; W632 (anti-MHC class 1)) and LAP following blockade with anti-αvβ6 antibody (10D5). Results are expressed relative to MMP-9 expression by VB6 cells on BSA-coated plastic (=100). Error bars represent standard deviation. These data suggest that αvβ6-LAP interaction modulates MMP-9 expression in VB6 cells.
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Related In: Results  -  Collection

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fig4: Binding of αvβ6 to LAP induces upregulation of MMP-9. Zymography for MMP-9. Cells were grown for 24 h in additive-free medium before supernatant sampling and cell counting. Samples containing equal volume per cell number were run on each gel with MMP-9 control. The intensity of the bands was measured by densitometric analysis and comparisons made within each gel to determine relative changes in MMP activity. (A) Zymogram showing upregulation of MMP-9 expression by VB6 cells when plated on fibronectin (FN) and LAP relative to BSA-coated plastic. Control is pro-MMP-9. (B) Zymogram showing the inhibition of MMP-9 expression by VB6 cells plated onto LAP following blockade with anti-αvβ6 antibody 10D5 relative to an irrelevant control antibody (Ctl; W632 (anti-MHC class 1)). (C) Densitometric analysis of zymograms from multiple separate experiments (n=4) showing MMP-9 expression by VB6 cells on BSA, LAP following incubation with an irrelevant control antibody (Ctl; W632 (anti-MHC class 1)) and LAP following blockade with anti-αvβ6 antibody (10D5). Results are expressed relative to MMP-9 expression by VB6 cells on BSA-coated plastic (=100). Error bars represent standard deviation. These data suggest that αvβ6-LAP interaction modulates MMP-9 expression in VB6 cells.
Mentions: Expression of MMP-9 on BSA-, fibronectin-, and LAP-coated tissue culture plastic was examined by zymography. Zymography on supernatant samples from cells grown in serum-free medium showed that the cells produced predominantly MMP-9 and that this enzyme was mainly present in proenzyme form. Similar to the effect observed on fibronectin (Thomas et al, 2001c), VB6 cells showed significant upregulation of MMP-9 when plated on LAP (P=<0.001; Figure 4A,CFigure 4

Bottom Line: Since TGF-beta1 is present in squamous carcinomas, it is possible that latency associated peptide may modulate malignant keratinocyte behaviour independently from the classical TGF-beta signalling pathways through its interaction with integrins.We show here that when latency associated peptide is immobilised onto a surface, it acts as an alpha(v)beta6-specific ligand for oral squamous carcinoma cells promoting adhesion and haptotactic migration in addition to alpha(v)beta6-dependent increase in pro-MMP-9 expression.In contrast, even very low concentrations of soluble latency associated peptide (0.1 microg ml(-1)) inhibited alpha(v)beta6-dependent adhesion, migration and invasion.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Pathology, Eastman Dental Institute, University College London, UK.

Show MeSH
Related in: MedlinePlus