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Creation of immortalised epithelial cells from ovarian endometrioma
BACKGROUND: Epithelial cells of endometriotic tissues are difficult to propagate in vitro as experimental material is scarce owing to their limited life span. However, there is an increasing concern regarding their malignant transformation in ovaries. The present study sought to generate their stabl...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3304406/ https://www.ncbi.nlm.nih.gov/pubmed/22353808 http://dx.doi.org/10.1038/bjc.2012.26 |
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author | Bono, Y Kyo, S Takakura, M Maida, Y Mizumoto, Y Nakamura, M Nomura, K Kiyono, T Inoue, M |
author_facet | Bono, Y Kyo, S Takakura, M Maida, Y Mizumoto, Y Nakamura, M Nomura, K Kiyono, T Inoue, M |
author_sort | Bono, Y |
collection | PubMed |
description | BACKGROUND: Epithelial cells of endometriotic tissues are difficult to propagate in vitro as experimental material is scarce owing to their limited life span. However, there is an increasing concern regarding their malignant transformation in ovaries. The present study sought to generate their stable culture system. METHODS AND RESULTS: Purified epithelial cells isolated from ovarian endometriomas using microscopic manipulation were successfully immortalised by combinatorial transfection of human cyclinD1, cdk4 and human telomerase reverse transcriptase (hTERT) genes, whereas the introduction of hTERT alone, or together with cdk4, was insufficient for immortalisation, leading to cellular senescence. We confirmed stable cytokeratin expression in the immortalised cells, proving their epithelial origin. These cells expressed progesterone receptor B and showed significant growth inhibition by various progestins. Oestrogen receptor (ER) expression was detected in these cells, albeit at low levels. Additional overexpression of ERα generated stable cells with oestrogen-dependent growth activation. Soft-agar colony formation assay and nude mice xenograft experiments demonstrated that these cells, even those with additional inactivation of p53, did not have transformed phenotypes. CONCLUSION: We for the first time generated immortalised epithelial cells from ovarian endometrioma that retained sex steroid responsiveness. These cells are invaluable tools not only for the consistent in vitro work but also for the study of molecular pathogenesis or carcinogenesis of endometriosis. |
format | Online Article Text |
id | pubmed-3304406 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-33044062013-03-13 Creation of immortalised epithelial cells from ovarian endometrioma Bono, Y Kyo, S Takakura, M Maida, Y Mizumoto, Y Nakamura, M Nomura, K Kiyono, T Inoue, M Br J Cancer Molecular Diagnostics BACKGROUND: Epithelial cells of endometriotic tissues are difficult to propagate in vitro as experimental material is scarce owing to their limited life span. However, there is an increasing concern regarding their malignant transformation in ovaries. The present study sought to generate their stable culture system. METHODS AND RESULTS: Purified epithelial cells isolated from ovarian endometriomas using microscopic manipulation were successfully immortalised by combinatorial transfection of human cyclinD1, cdk4 and human telomerase reverse transcriptase (hTERT) genes, whereas the introduction of hTERT alone, or together with cdk4, was insufficient for immortalisation, leading to cellular senescence. We confirmed stable cytokeratin expression in the immortalised cells, proving their epithelial origin. These cells expressed progesterone receptor B and showed significant growth inhibition by various progestins. Oestrogen receptor (ER) expression was detected in these cells, albeit at low levels. Additional overexpression of ERα generated stable cells with oestrogen-dependent growth activation. Soft-agar colony formation assay and nude mice xenograft experiments demonstrated that these cells, even those with additional inactivation of p53, did not have transformed phenotypes. CONCLUSION: We for the first time generated immortalised epithelial cells from ovarian endometrioma that retained sex steroid responsiveness. These cells are invaluable tools not only for the consistent in vitro work but also for the study of molecular pathogenesis or carcinogenesis of endometriosis. Nature Publishing Group 2012-03-13 2012-02-21 /pmc/articles/PMC3304406/ /pubmed/22353808 http://dx.doi.org/10.1038/bjc.2012.26 Text en Copyright © 2012 Cancer Research UK https://creativecommons.org/licenses/by/4.0/This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material.If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Molecular Diagnostics Bono, Y Kyo, S Takakura, M Maida, Y Mizumoto, Y Nakamura, M Nomura, K Kiyono, T Inoue, M Creation of immortalised epithelial cells from ovarian endometrioma |
title | Creation of immortalised epithelial cells from ovarian endometrioma |
title_full | Creation of immortalised epithelial cells from ovarian endometrioma |
title_fullStr | Creation of immortalised epithelial cells from ovarian endometrioma |
title_full_unstemmed | Creation of immortalised epithelial cells from ovarian endometrioma |
title_short | Creation of immortalised epithelial cells from ovarian endometrioma |
title_sort | creation of immortalised epithelial cells from ovarian endometrioma |
topic | Molecular Diagnostics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3304406/ https://www.ncbi.nlm.nih.gov/pubmed/22353808 http://dx.doi.org/10.1038/bjc.2012.26 |
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