Successful immortalization of endometrial glandular cells with normal structural and functional characteristics
Successful immortalization of endometrial glandular cells with normal structural and functional characteristics
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DOI：
10.1016/s0002-9440(10)63583-3
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发表时间：
2003-12-01
影响因子：
6
通讯作者：
Inoue, M
中科院分区：
文献类型：
--
作者：
Kyo, S;Nakamura, M;Inoue, M
人的子宫内膜是一个动态的组织，其增殖活动在整个月经周期中发生巨大变化，受性类固醇激素的精确调节。在塑料培养皿中培养的原代子宫内膜上皮细胞在2周内衰老，对子宫内膜生物学的更全面的了解被推迟，部分原因是缺乏子宫内膜上皮细胞的体外培养模型。我们的目标是建立永生化的人类子宫内膜腺细胞，以保留原始细胞的正常功能和特征。由于Rb/p16和P53通路被认为是早期传代的上皮衰老的关键元件，我们使用人乳头瘤病毒E6/E7来靶向这些通路。人类乳头瘤病毒16E6/E7的表达和端粒酶的激活相结合，通过人端粒酶逆转录酶(HTERT)的引入，成功地使子宫内膜腺细胞永生化。仅E6/E7的表达就足以延长其寿命超过20个群体倍增，但进一步需要端粒酶激活才能使细胞在40个群体倍增时经历随后的复制衰老。分离的永生化细胞不含染色体异常或仅含非克隆。畸形，保留了对性类固醇激素的反应性，在三维培养上表现为腺体结构，在软琼脂或裸鼠体内缺乏转化的表型。这些发现支持Rb失活/P53失活和端粒酶激活是子宫内膜上皮细胞永生化所必需的，但子宫内膜癌的发生还需要其他因素。我们已建立的细胞系在激素功能、子宫内膜生物学和子宫内膜癌变的研究中显示出巨大的希望。
The human endometrium is a dynamic tissue, the proliferative activity of which dramatically changes throughout the menstrual cycle, with exquisite regulation by sex-steroid hormones. Primary endometrial epithelial cells fall into senescence within 2 weeks when cultured on plastic dishes, and more complete understanding of endometrial biology has been delayed because of, in part, a lack of an in vitro culture model for endometrial epithelial cells. our goal was to establish immortalized human endometrial glandular cells that retain the normal functions and characteristics of the primary cells. Because the Rb/p16 and P53 pathways are known to be critical elements of epithelial senescence in early passages, we used human papillomavirus E6/E7 to target these pathways. The combination of human papillomavirus-16 E6/E7 expression and telomerase activation by the introduction of human telomerase reverse transcriptase (hTERT) led to successful immortalization of the endometrial glandular cells. E6/E7 expression alone was sufficient to extend their fife span more than 20 population doublings, but the telomerase activation was further required to enable the cells to pass through the subsequent replicative senescence at 40 population doublings. Isolated immortalized cells contained no chromosomal abnormalities or only nonclonal. aberrations, retained responsiveness to sex-steroid hormones, exhibited glandular structure on three-dimensional culture, and lacked transformed phenotypes on soft agar or in nude mice. These findings support the notion that both Rb inactivation/p53 inactivation and telomerase activation are necessary to immortalize endometrial epithelial cells, but additional factors are required for endometrial carcinogenesis. our established cell lines show great promise for investigation of hormone functions, endometrial biology, and endometrial carcinogenesis.