Creation of immortalised epithelial cells from ovarian endometrioma.
Creation of immortalised epithelial cells from ovarian endometrioma.
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DOI：
10.1038/bjc.2012.26
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发表时间：
2012-03-13
影响因子：
8.8
通讯作者：
Inoue, M.
中科院分区：
文献类型：
--
作者：
Bono, Y.;Kyo, S.;Takakura, M.;Maida, Y.;Mizumoto, Y.;Nakamura, M.;Nomura, K.;Kiyono, T.;Inoue, M.
子宫内膜异位组织的上皮细胞寿命有限，实验材料匮乏，难以在体外增殖。然而，人们越来越担心卵巢的恶变。本研究旨在建立其稳定的培养系统。通过显微操作从卵巢子宫内膜异位瘤中分离纯化的上皮细胞，通过人细胞周期蛋白D1、cdk4和人端粒酶逆转录酶（hTERT）基因的组合转染，成功地永生化，而单独引入hTERT或与cdk4一起引入，不足以实现永生化，导致细胞衰老。我们证实了永生化细胞中稳定的细胞角蛋白表达，证明了它们的上皮起源。这些细胞表达孕酮受体 B，并显示出各种孕激素的显着生长抑制作用。在这些细胞中检测到雌激素受体（ER）表达，尽管水平较低。 ERα 的额外过度表达产生了具有雌激素依赖性生长激活的稳定细胞。软琼脂集落形成测定和裸鼠异种移植实验表明，这些细胞，即使是那些 p53 额外失活的细胞，也没有转化的表型。我们首次从卵巢子宫内膜瘤中产生了永生化的上皮细胞，该细胞保留了性类固醇的反应性。这些细胞不仅对于一致的体外工作而且对于子宫内膜异位症的分子发病机制或癌发生的研究都是宝贵的工具。
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. 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. 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.
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影响因子：
3.6
作者：
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通讯作者：
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Inoue, M
DOI：
10.1016/s0301-2115(03)00047-2
发表时间：
2003-07-01
影响因子：
2.6
作者：
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通讯作者：
Kosebay, D