Loss of MMP-8 in ductal carcinoma in situ (DCIS)-associated myoepithelial cells contributes to tumour promotion through altered adhesive and proteolytic function

<h3>Background</h3><p dir="ltr">Normal myoepithelial cells (MECs) play an important tumour-suppressor role in the breast but display an altered phenotype in ductal carcinoma in situ (DCIS), gaining tumour-promoter functions. Matrix metalloproteinase-8 (MMP-8) is expressed...

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التفاصيل البيبلوغرافية
المؤلف الرئيسي: Muge Sarper (3855454) (author)
مؤلفون آخرون: Michael D. Allen (263658) (author), Jenny Gomm (3855451) (author), Linda Haywood (3855460) (author), Julie Decock (44558) (author), Sally Thirkettle (702179) (author), Ahsen Ustaoglu (3855448) (author), Shah-Jalal Sarker (3502589) (author), John Marshall (39501) (author), Dylan R. Edwards (7957886) (author), J. Louise Jones (7687964) (author)
منشور في: 2017
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author Muge Sarper (3855454)
author2 Michael D. Allen (263658)
Jenny Gomm (3855451)
Linda Haywood (3855460)
Julie Decock (44558)
Sally Thirkettle (702179)
Ahsen Ustaoglu (3855448)
Shah-Jalal Sarker (3502589)
John Marshall (39501)
Dylan R. Edwards (7957886)
J. Louise Jones (7687964)
author2_role author
author
author
author
author
author
author
author
author
author
author_facet Muge Sarper (3855454)
Michael D. Allen (263658)
Jenny Gomm (3855451)
Linda Haywood (3855460)
Julie Decock (44558)
Sally Thirkettle (702179)
Ahsen Ustaoglu (3855448)
Shah-Jalal Sarker (3502589)
John Marshall (39501)
Dylan R. Edwards (7957886)
J. Louise Jones (7687964)
author_role author
dc.creator.none.fl_str_mv Muge Sarper (3855454)
Michael D. Allen (263658)
Jenny Gomm (3855451)
Linda Haywood (3855460)
Julie Decock (44558)
Sally Thirkettle (702179)
Ahsen Ustaoglu (3855448)
Shah-Jalal Sarker (3502589)
John Marshall (39501)
Dylan R. Edwards (7957886)
J. Louise Jones (7687964)
dc.date.none.fl_str_mv 2017-03-23T03:00:00Z
dc.identifier.none.fl_str_mv 10.1186/s13058-017-0822-9
dc.relation.none.fl_str_mv https://figshare.com/articles/journal_contribution/Loss_of_MMP-8_in_ductal_carcinoma_in_situ_DCIS_-associated_myoepithelial_cells_contributes_to_tumour_promotion_through_altered_adhesive_and_proteolytic_function/27088003
dc.rights.none.fl_str_mv CC BY 4.0
info:eu-repo/semantics/openAccess
dc.subject.none.fl_str_mv Biomedical and clinical sciences
Oncology and carcinogenesis
Ductal carcinoma in situ
Myoepithelial cell
Microenvironment
MMP-8
Adhesion
Hemidesmosomes
Organotypic assays
Invasion
dc.title.none.fl_str_mv Loss of MMP-8 in ductal carcinoma in situ (DCIS)-associated myoepithelial cells contributes to tumour promotion through altered adhesive and proteolytic function
dc.type.none.fl_str_mv Text
Journal contribution
info:eu-repo/semantics/publishedVersion
text
contribution to journal
description <h3>Background</h3><p dir="ltr">Normal myoepithelial cells (MECs) play an important tumour-suppressor role in the breast but display an altered phenotype in ductal carcinoma in situ (DCIS), gaining tumour-promoter functions. Matrix metalloproteinase-8 (MMP-8) is expressed by normal MECs but is lost in DCIS. This study investigated the function of MMP-8 in MECs and the impact of its loss in DCIS.</p><h3>Methods</h3><p dir="ltr">Primary normal and DCIS-associated MECs, and normal (N-1089) and DCIS-modified myoepithelial (β6-1089) cell lines, were used to assess MMP-8 expression and function. β6-1089 lacking MMP-8 were transfected with MMP-8 WT and catalytically inactive MMP-8 EA, and MMP-8 in N-1089 MEC was knocked down with siRNA. The effect on adhesion and migration to extracellular matrix (ECM), localisation of α6β4 integrin to hemidesmosomes (HD), TGF-β signalling and gelatinase activity was measured. The effect of altering MEC MMP-8 expression on tumour cell invasion was investigated in 2D and 3D organotypic models.</p><h3>Results</h3><p dir="ltr">Assessment of primary cells and MEC lines confirmed expression of MMP-8 in normal MEC and its loss in DCIS-MEC. Over-expression of MMP-8 WT but not MMP-8 EA in β6-1089 cells increased adhesion to ECM proteins and reduced migration. Conversely, knock-down of MMP-8 in N-1089 reduced adhesion and increased migration. Expression of MMP-8 WT in β6-1089 led to greater localisation of α6β4 to HD and reduced retraction fibre formation, this being reversed by MMP-8 knock-down in N-1089. Over-expression of MMP-8 WT reduced TGF-β signalling and gelatinolytic activity. MMP-8 knock-down enhanced TGF-β signalling and gelatinolytic activity, which was reversed by blocking MMP-9 by knock-down or an inhibitor. MMP-8 WT but not MMP-8 EA over-expression in β6-1089 reduced breast cancer cell invasion in 2D and 3D invasion assays, while MMP-8 knock-down in N-1089 enhanced cancer cell invasion. Staining of breast cancer cases for MMP-8 revealed a statistically significant loss of MMP-8 expression in DCIS with invasion versus pure DCIS (<i>p</i> = 0.001).</p><h3>Conclusions</h3><p dir="ltr">These data indicate MMP-8 is a vital component of the myoepithelial tumour-suppressor function. It restores MEC interaction with the matrix, opposes TGF-β signalling and MMP-9 proteolysis, which contributes to inhibition of tumour cell invasion. Assessment of MMP-8 expression may help to determine risk of DCIS progression.</p><h2>Other Information</h2><p dir="ltr">Published in: Breast Cancer Research<br>License: <a href="https://creativecommons.org/licenses/by/4.0/deed.en" rel="noreferrer noopener" target="_blank">https://creativecommons.org/licenses/by/4.0/</a>  <br>See article on publisher's website: <a href="https://dx.doi.org/10.1186/s13058-017-0822-9" target="_blank">https://dx.doi.org/10.1186/s13058-017-0822-9</a></p>
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identifier_str_mv 10.1186/s13058-017-0822-9
network_acronym_str Manara2
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oai_identifier_str oai:figshare.com:article/27088003
publishDate 2017
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rights_invalid_str_mv CC BY 4.0
spelling Loss of MMP-8 in ductal carcinoma in situ (DCIS)-associated myoepithelial cells contributes to tumour promotion through altered adhesive and proteolytic functionMuge Sarper (3855454)Michael D. Allen (263658)Jenny Gomm (3855451)Linda Haywood (3855460)Julie Decock (44558)Sally Thirkettle (702179)Ahsen Ustaoglu (3855448)Shah-Jalal Sarker (3502589)John Marshall (39501)Dylan R. Edwards (7957886)J. Louise Jones (7687964)Biomedical and clinical sciencesOncology and carcinogenesisDuctal carcinoma in situMyoepithelial cellMicroenvironmentMMP-8AdhesionHemidesmosomesOrganotypic assaysInvasion<h3>Background</h3><p dir="ltr">Normal myoepithelial cells (MECs) play an important tumour-suppressor role in the breast but display an altered phenotype in ductal carcinoma in situ (DCIS), gaining tumour-promoter functions. Matrix metalloproteinase-8 (MMP-8) is expressed by normal MECs but is lost in DCIS. This study investigated the function of MMP-8 in MECs and the impact of its loss in DCIS.</p><h3>Methods</h3><p dir="ltr">Primary normal and DCIS-associated MECs, and normal (N-1089) and DCIS-modified myoepithelial (β6-1089) cell lines, were used to assess MMP-8 expression and function. β6-1089 lacking MMP-8 were transfected with MMP-8 WT and catalytically inactive MMP-8 EA, and MMP-8 in N-1089 MEC was knocked down with siRNA. The effect on adhesion and migration to extracellular matrix (ECM), localisation of α6β4 integrin to hemidesmosomes (HD), TGF-β signalling and gelatinase activity was measured. The effect of altering MEC MMP-8 expression on tumour cell invasion was investigated in 2D and 3D organotypic models.</p><h3>Results</h3><p dir="ltr">Assessment of primary cells and MEC lines confirmed expression of MMP-8 in normal MEC and its loss in DCIS-MEC. Over-expression of MMP-8 WT but not MMP-8 EA in β6-1089 cells increased adhesion to ECM proteins and reduced migration. Conversely, knock-down of MMP-8 in N-1089 reduced adhesion and increased migration. Expression of MMP-8 WT in β6-1089 led to greater localisation of α6β4 to HD and reduced retraction fibre formation, this being reversed by MMP-8 knock-down in N-1089. Over-expression of MMP-8 WT reduced TGF-β signalling and gelatinolytic activity. MMP-8 knock-down enhanced TGF-β signalling and gelatinolytic activity, which was reversed by blocking MMP-9 by knock-down or an inhibitor. MMP-8 WT but not MMP-8 EA over-expression in β6-1089 reduced breast cancer cell invasion in 2D and 3D invasion assays, while MMP-8 knock-down in N-1089 enhanced cancer cell invasion. Staining of breast cancer cases for MMP-8 revealed a statistically significant loss of MMP-8 expression in DCIS with invasion versus pure DCIS (<i>p</i> = 0.001).</p><h3>Conclusions</h3><p dir="ltr">These data indicate MMP-8 is a vital component of the myoepithelial tumour-suppressor function. It restores MEC interaction with the matrix, opposes TGF-β signalling and MMP-9 proteolysis, which contributes to inhibition of tumour cell invasion. Assessment of MMP-8 expression may help to determine risk of DCIS progression.</p><h2>Other Information</h2><p dir="ltr">Published in: Breast Cancer Research<br>License: <a href="https://creativecommons.org/licenses/by/4.0/deed.en" rel="noreferrer noopener" target="_blank">https://creativecommons.org/licenses/by/4.0/</a>  <br>See article on publisher's website: <a href="https://dx.doi.org/10.1186/s13058-017-0822-9" target="_blank">https://dx.doi.org/10.1186/s13058-017-0822-9</a></p>2017-03-23T03:00:00ZTextJournal contributioninfo:eu-repo/semantics/publishedVersiontextcontribution to journal10.1186/s13058-017-0822-9https://figshare.com/articles/journal_contribution/Loss_of_MMP-8_in_ductal_carcinoma_in_situ_DCIS_-associated_myoepithelial_cells_contributes_to_tumour_promotion_through_altered_adhesive_and_proteolytic_function/27088003CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/270880032017-03-23T03:00:00Z
spellingShingle Loss of MMP-8 in ductal carcinoma in situ (DCIS)-associated myoepithelial cells contributes to tumour promotion through altered adhesive and proteolytic function
Muge Sarper (3855454)
Biomedical and clinical sciences
Oncology and carcinogenesis
Ductal carcinoma in situ
Myoepithelial cell
Microenvironment
MMP-8
Adhesion
Hemidesmosomes
Organotypic assays
Invasion
status_str publishedVersion
title Loss of MMP-8 in ductal carcinoma in situ (DCIS)-associated myoepithelial cells contributes to tumour promotion through altered adhesive and proteolytic function
title_full Loss of MMP-8 in ductal carcinoma in situ (DCIS)-associated myoepithelial cells contributes to tumour promotion through altered adhesive and proteolytic function
title_fullStr Loss of MMP-8 in ductal carcinoma in situ (DCIS)-associated myoepithelial cells contributes to tumour promotion through altered adhesive and proteolytic function
title_full_unstemmed Loss of MMP-8 in ductal carcinoma in situ (DCIS)-associated myoepithelial cells contributes to tumour promotion through altered adhesive and proteolytic function
title_short Loss of MMP-8 in ductal carcinoma in situ (DCIS)-associated myoepithelial cells contributes to tumour promotion through altered adhesive and proteolytic function
title_sort Loss of MMP-8 in ductal carcinoma in situ (DCIS)-associated myoepithelial cells contributes to tumour promotion through altered adhesive and proteolytic function
topic Biomedical and clinical sciences
Oncology and carcinogenesis
Ductal carcinoma in situ
Myoepithelial cell
Microenvironment
MMP-8
Adhesion
Hemidesmosomes
Organotypic assays
Invasion