<it>In vitro </it>and <it>in vivo </it>MMP gene expression localisation by <it>In Situ</it>-RT-PCR in cell culture and paraffin embedded human breast cancer cell line xenografts
<p>Abstract</p> <p>Background</p> <p>Members of the matrix metalloproteinase (MMP) family of proteases are required for the degradation of the basement membrane and extracellular matrix in both normal and pathological conditions. <it>In vitro</it>, MT1-MMP (...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2006-01-01
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| Series: | BMC Cancer |
| Online Access: | http://www.biomedcentral.com/1471-2407/6/18 |
| _version_ | 1828235803094941696 |
|---|---|
| author | Irving Michael G Trezise Ann EO Irving Rachel E Thompson Erik W Haupt Larisa M Griffiths Lyn R |
| author_facet | Irving Michael G Trezise Ann EO Irving Rachel E Thompson Erik W Haupt Larisa M Griffiths Lyn R |
| author_sort | Irving Michael G |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Members of the matrix metalloproteinase (MMP) family of proteases are required for the degradation of the basement membrane and extracellular matrix in both normal and pathological conditions. <it>In vitro</it>, MT1-MMP (MMP-14, membrane type-1-MMP) expression is higher in more invasive human breast cancer (HBC) cell lines, whilst <it>in vivo </it>its expression has been associated with the stroma surrounding breast tumours. MMP-1 (interstitial collagenase) has been associated with MDA-MB-231 invasion <it>in vitro</it>, while MMP-3 (stromelysin-1) has been localised around invasive cells of breast tumours <it>in vivo</it>. As MMPs are not stored intracellularly, the ability to localise their expression to their cells of origin is difficult.</p> <p>Methods</p> <p>We utilised the unique <it>in situ</it>-reverse transcription-polymerase chain reaction (<it>IS</it>-RT-PCR) methodology to localise the <it>in vitro </it>and <it>in vivo </it>gene expression of MT1-MMP, MMP-1 and MMP-3 in human breast cancer. <it>In vitro</it>, MMP induction was examined in the MDA-MB-231 and MCF-7 HBC cell lines following exposure to Concanavalin A (Con A). <it>In vivo</it>, we examined their expression in archival paraffin embedded xenografts derived from a range of HBC cell lines of varied invasive and metastatic potential. Mouse xenografts are heterogenous, containing neoplastic human parenchyma with mouse stroma and vasculature and provide a reproducible <it>in vivo </it>model system correlated to the human disease state.</p> <p>Results</p> <p><it>In vitro</it>, exposure to Con A increased MT1-MMP gene expression in MDA-MB-231 cells and decreased MT1-MMP gene expression in MCF-7 cells. MMP-1 and MMP-3 gene expression remained unchanged in both cell lines. <it>In vivo</it>, stromal cells recruited into each xenograft demonstrated differences in localised levels of MMP gene expression. Specifically, MDA-MB-231, MDA-MB-435 and Hs578T HBC cell lines are able to influence MMP gene expression in the surrounding stroma.</p> <p>Conclusion</p> <p>We have demonstrated the applicability and sensitivity of <it>IS</it>-RT-PCR for the examination of MMP gene expression both <it>in vitro </it>and <it>in vivo</it>. Induction of MMP gene expression in both the epithelial tumour cells and surrounding stromal cells is associated with increased metastatic potential. Our data demonstrate the contribution of the stroma to epithelial MMP gene expression, and highlight the complexity of the role of MMPs in the stromal-epithelial interactions within breast carcinoma.</p> |
| first_indexed | 2024-04-12T20:22:02Z |
| format | Article |
| id | doaj.art-d295c41d10aa442881cc38cc9fcd34f8 |
| institution | Directory Open Access Journal |
| issn | 1471-2407 |
| language | English |
| last_indexed | 2024-04-12T20:22:02Z |
| publishDate | 2006-01-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Cancer |
| spelling | doaj.art-d295c41d10aa442881cc38cc9fcd34f82022-12-22T03:17:59ZengBMCBMC Cancer1471-24072006-01-01611810.1186/1471-2407-6-18<it>In vitro </it>and <it>in vivo </it>MMP gene expression localisation by <it>In Situ</it>-RT-PCR in cell culture and paraffin embedded human breast cancer cell line xenograftsIrving Michael GTrezise Ann EOIrving Rachel EThompson Erik WHaupt Larisa MGriffiths Lyn R<p>Abstract</p> <p>Background</p> <p>Members of the matrix metalloproteinase (MMP) family of proteases are required for the degradation of the basement membrane and extracellular matrix in both normal and pathological conditions. <it>In vitro</it>, MT1-MMP (MMP-14, membrane type-1-MMP) expression is higher in more invasive human breast cancer (HBC) cell lines, whilst <it>in vivo </it>its expression has been associated with the stroma surrounding breast tumours. MMP-1 (interstitial collagenase) has been associated with MDA-MB-231 invasion <it>in vitro</it>, while MMP-3 (stromelysin-1) has been localised around invasive cells of breast tumours <it>in vivo</it>. As MMPs are not stored intracellularly, the ability to localise their expression to their cells of origin is difficult.</p> <p>Methods</p> <p>We utilised the unique <it>in situ</it>-reverse transcription-polymerase chain reaction (<it>IS</it>-RT-PCR) methodology to localise the <it>in vitro </it>and <it>in vivo </it>gene expression of MT1-MMP, MMP-1 and MMP-3 in human breast cancer. <it>In vitro</it>, MMP induction was examined in the MDA-MB-231 and MCF-7 HBC cell lines following exposure to Concanavalin A (Con A). <it>In vivo</it>, we examined their expression in archival paraffin embedded xenografts derived from a range of HBC cell lines of varied invasive and metastatic potential. Mouse xenografts are heterogenous, containing neoplastic human parenchyma with mouse stroma and vasculature and provide a reproducible <it>in vivo </it>model system correlated to the human disease state.</p> <p>Results</p> <p><it>In vitro</it>, exposure to Con A increased MT1-MMP gene expression in MDA-MB-231 cells and decreased MT1-MMP gene expression in MCF-7 cells. MMP-1 and MMP-3 gene expression remained unchanged in both cell lines. <it>In vivo</it>, stromal cells recruited into each xenograft demonstrated differences in localised levels of MMP gene expression. Specifically, MDA-MB-231, MDA-MB-435 and Hs578T HBC cell lines are able to influence MMP gene expression in the surrounding stroma.</p> <p>Conclusion</p> <p>We have demonstrated the applicability and sensitivity of <it>IS</it>-RT-PCR for the examination of MMP gene expression both <it>in vitro </it>and <it>in vivo</it>. Induction of MMP gene expression in both the epithelial tumour cells and surrounding stromal cells is associated with increased metastatic potential. Our data demonstrate the contribution of the stroma to epithelial MMP gene expression, and highlight the complexity of the role of MMPs in the stromal-epithelial interactions within breast carcinoma.</p>http://www.biomedcentral.com/1471-2407/6/18 |
| spellingShingle | Irving Michael G Trezise Ann EO Irving Rachel E Thompson Erik W Haupt Larisa M Griffiths Lyn R <it>In vitro </it>and <it>in vivo </it>MMP gene expression localisation by <it>In Situ</it>-RT-PCR in cell culture and paraffin embedded human breast cancer cell line xenografts BMC Cancer |
| title | <it>In vitro </it>and <it>in vivo </it>MMP gene expression localisation by <it>In Situ</it>-RT-PCR in cell culture and paraffin embedded human breast cancer cell line xenografts |
| title_full | <it>In vitro </it>and <it>in vivo </it>MMP gene expression localisation by <it>In Situ</it>-RT-PCR in cell culture and paraffin embedded human breast cancer cell line xenografts |
| title_fullStr | <it>In vitro </it>and <it>in vivo </it>MMP gene expression localisation by <it>In Situ</it>-RT-PCR in cell culture and paraffin embedded human breast cancer cell line xenografts |
| title_full_unstemmed | <it>In vitro </it>and <it>in vivo </it>MMP gene expression localisation by <it>In Situ</it>-RT-PCR in cell culture and paraffin embedded human breast cancer cell line xenografts |
| title_short | <it>In vitro </it>and <it>in vivo </it>MMP gene expression localisation by <it>In Situ</it>-RT-PCR in cell culture and paraffin embedded human breast cancer cell line xenografts |
| title_sort | it in vitro it and it in vivo it mmp gene expression localisation by it in situ it rt pcr in cell culture and paraffin embedded human breast cancer cell line xenografts |
| url | http://www.biomedcentral.com/1471-2407/6/18 |
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