Creation of immortalised epithelial cells from ovarian endometrioma Br J Cancer

Yukiko Bono, Satoru Kyo, Masahiro Takakura, Yoshiko Maida, Yasunari Mizumoto, Manabu Nakamura, Kenta Nomura, Toru Kiyono, M. Inoue
2013 · vol. 122(2) , pp. 37–38 · W2783491121
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This study successfully immortalized epithelial cells from ovarian endometriomas by transfecting human cyclin D1, cdk4, and hTERT genes, enabling stable culture and characterization of their responsiveness to sex steroids.

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The paper studied how to establish a stable, expandable in vitro epithelial cell culture system from ovarian endometrioma tissue, where native epithelial cells are difficult to propagate due to limited lifespan. Purified epithelial cells were isolated by microscopic manipulation and immortalised using combinatorial transfection of cyclin D1, CDK4, and hTERT, while hTERT alone or hTERT with CDK4 was insufficient and led to senescence. The resulting immortalised cells retained epithelial cytokeratin expression and progesterone receptor B, exhibited progesterone- and progestin-related growth inhibition, expressed estrogen receptor at low levels, and after ERα overexpression showed estrogen-dependent growth activation. Soft-agar assays and nude mice xenograft experiments showed no transformed phenotype, including in cells with p53 inactivation, and the authors acknowledge the approach as a tool for studying endometriosis pathogenesis/carcinogenesis. This paper is centrally about endometriosis — it reports creation of immortalised epithelial cells specifically derived from ovarian endometrioma that preserve sex steroid responsiveness.

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Creation of immortalised epithelial cells from ovarian endometrioma Bibliographic Information - Other Title - - 子宮内膜症性卵巣嚢胞から分離した子宮内膜症上皮不死化細胞株の樹立 - Published - 2013-06-01 - Resource Type - departmental bulletin paper - Publisher - 2013-06-01 Search this article 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. © 2012 Cancer Research UK All rights reserved. Journal - - 金沢大学十全医学会雑誌 = Journal of the Juzen Medical Society - 金沢大学十全医学会雑誌 = Journal of the Juzen Medical Society 122 (2), 37-38, 2013-06-01 2013-06-01 Details 詳細情報について - - CRID - 1050001202729724032 - - NII Article ID - 10031193194 - - NII Book ID - AN00044397 - - ISSN - 00227226 - - HANDLE - 2297/35533 - - Text Lang - ja - - Article Type - departmental bulletin paper - - Data Source - - IRDB - CiNii Articles

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