CD73 complexes with emmprin to regulate MMP-2 production from co-cultured sarcoma cells and fibroblasts
Abstract Background Interaction between cancer cells and fibroblasts mediated by extracellular matrix metalloproteinase inducer (emmprin, CD147) is important in the invasion and proliferation of cancer cells. However, the exact mechanism of emmprin mediated stimulation of matrix metalloprotease-2 (M...
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| Format: | Article |
| Language: | English |
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BMC
2019-09-01
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| Series: | BMC Cancer |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12885-019-6127-x |
| _version_ | 1818934527412666368 |
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| author | M. Aoki K. Koga M. Miyazaki M. Hamasaki N. Koshikawa M. Oyama H. Kozuka-Hata M. Seiki B. P. Toole K. Nabeshima |
| author_facet | M. Aoki K. Koga M. Miyazaki M. Hamasaki N. Koshikawa M. Oyama H. Kozuka-Hata M. Seiki B. P. Toole K. Nabeshima |
| author_sort | M. Aoki |
| collection | DOAJ |
| description | Abstract Background Interaction between cancer cells and fibroblasts mediated by extracellular matrix metalloproteinase inducer (emmprin, CD147) is important in the invasion and proliferation of cancer cells. However, the exact mechanism of emmprin mediated stimulation of matrix metalloprotease-2 (MMP-2) production from fibroblasts has not been elucidated. Our previous studies using an inhibitory peptide against emmprin suggested the presence of a molecule on the cell membrane which forms a complex with emmprin. Here we show that CD73 expressed on fibroblasts interacts with emmprin and is a required factor for MMP-2 production in co-cultures of sarcoma cells with fibroblasts. Methods CD73 along with CD99 was identified by mass spectrometry analysis as an emmprin interacting molecule from a co-culture of cancer cells (epithelioid sarcoma cell line FU-EPS-1) and fibroblasts (immortalized fibroblasts cell line ST353i). MMP-2 production was measured by immunoblot and ELISA. The formation of complexes of CD73 with emmprin was confirmed by immunoprecipitation, and their co-localization in tumor cells and fibroblasts was shown by fluorescent immunostaining and proximity ligation assays. Results Stimulated MMP-2 production in co-culture of cancer cells and fibroblasts was completely suppressed by siRNA knockdown of CD73, but not by CD99 knockdown. MMP-2 production was not suppressed by CD73-specific enzyme inhibitor (APCP). However, MMP-2 production was decreased by CD73 neutralizing antibodies, suggesting that CD73-mediated suppression of MMP-2 production is non-enzymatic. In human epithelioid sarcoma tissues, emmprin was immunohistochemically detected to be mainly expressed in tumor cells, and CD73 was expressed in fibroblasts and tumor cells: emmprin and CD73 were co-localized predominantly on tumor cells. Conclusion This study provides a novel insight into the role of CD73 in emmprin-mediated regulation of MMP-2 production. |
| first_indexed | 2024-12-20T05:05:41Z |
| format | Article |
| id | doaj.art-32723670cf54494ba644efc191798fa0 |
| institution | Directory Open Access Journal |
| issn | 1471-2407 |
| language | English |
| last_indexed | 2024-12-20T05:05:41Z |
| publishDate | 2019-09-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Cancer |
| spelling | doaj.art-32723670cf54494ba644efc191798fa02022-12-21T19:52:24ZengBMCBMC Cancer1471-24072019-09-0119111310.1186/s12885-019-6127-xCD73 complexes with emmprin to regulate MMP-2 production from co-cultured sarcoma cells and fibroblastsM. Aoki0K. Koga1M. Miyazaki2M. Hamasaki3N. Koshikawa4M. Oyama5H. Kozuka-Hata6M. Seiki7B. P. Toole8K. Nabeshima9Department of Pathology, Fukuoka University School of MedicineDepartment of Pathology, Fukuoka University School of MedicineDepartment of Pathology, Fukuoka University School of MedicineDepartment of Pathology, Fukuoka University School of MedicineDivision of Cancer Cell Research, Kanagawa Cancer Center Research InstituteMedical Proteomics Laboratory, Institute of Medical Science, University of TokyoMedical Proteomics Laboratory, Institute of Medical Science, University of TokyoDivision of Cancer Cell Research, Institute of Medical Science, University of TokyoRegenerative Medicine & Cell Biology, Medical University of South CarolinaDepartment of Pathology, Fukuoka University School of MedicineAbstract Background Interaction between cancer cells and fibroblasts mediated by extracellular matrix metalloproteinase inducer (emmprin, CD147) is important in the invasion and proliferation of cancer cells. However, the exact mechanism of emmprin mediated stimulation of matrix metalloprotease-2 (MMP-2) production from fibroblasts has not been elucidated. Our previous studies using an inhibitory peptide against emmprin suggested the presence of a molecule on the cell membrane which forms a complex with emmprin. Here we show that CD73 expressed on fibroblasts interacts with emmprin and is a required factor for MMP-2 production in co-cultures of sarcoma cells with fibroblasts. Methods CD73 along with CD99 was identified by mass spectrometry analysis as an emmprin interacting molecule from a co-culture of cancer cells (epithelioid sarcoma cell line FU-EPS-1) and fibroblasts (immortalized fibroblasts cell line ST353i). MMP-2 production was measured by immunoblot and ELISA. The formation of complexes of CD73 with emmprin was confirmed by immunoprecipitation, and their co-localization in tumor cells and fibroblasts was shown by fluorescent immunostaining and proximity ligation assays. Results Stimulated MMP-2 production in co-culture of cancer cells and fibroblasts was completely suppressed by siRNA knockdown of CD73, but not by CD99 knockdown. MMP-2 production was not suppressed by CD73-specific enzyme inhibitor (APCP). However, MMP-2 production was decreased by CD73 neutralizing antibodies, suggesting that CD73-mediated suppression of MMP-2 production is non-enzymatic. In human epithelioid sarcoma tissues, emmprin was immunohistochemically detected to be mainly expressed in tumor cells, and CD73 was expressed in fibroblasts and tumor cells: emmprin and CD73 were co-localized predominantly on tumor cells. Conclusion This study provides a novel insight into the role of CD73 in emmprin-mediated regulation of MMP-2 production.http://link.springer.com/article/10.1186/s12885-019-6127-xEmmprinCD73MMP-2SarcomaFibroblasts |
| spellingShingle | M. Aoki K. Koga M. Miyazaki M. Hamasaki N. Koshikawa M. Oyama H. Kozuka-Hata M. Seiki B. P. Toole K. Nabeshima CD73 complexes with emmprin to regulate MMP-2 production from co-cultured sarcoma cells and fibroblasts BMC Cancer Emmprin CD73 MMP-2 Sarcoma Fibroblasts |
| title | CD73 complexes with emmprin to regulate MMP-2 production from co-cultured sarcoma cells and fibroblasts |
| title_full | CD73 complexes with emmprin to regulate MMP-2 production from co-cultured sarcoma cells and fibroblasts |
| title_fullStr | CD73 complexes with emmprin to regulate MMP-2 production from co-cultured sarcoma cells and fibroblasts |
| title_full_unstemmed | CD73 complexes with emmprin to regulate MMP-2 production from co-cultured sarcoma cells and fibroblasts |
| title_short | CD73 complexes with emmprin to regulate MMP-2 production from co-cultured sarcoma cells and fibroblasts |
| title_sort | cd73 complexes with emmprin to regulate mmp 2 production from co cultured sarcoma cells and fibroblasts |
| topic | Emmprin CD73 MMP-2 Sarcoma Fibroblasts |
| url | http://link.springer.com/article/10.1186/s12885-019-6127-x |
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