Transformation of Human Endometrial Epithelial Cells

人子宫内膜上皮细胞的转化

• 批准号: 6709781
• 负责人: David G. Kaufman
• 金额: $ 13.14万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-09 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6709781
• 关键词: basement membrane; carcinogenesis; cell proliferation; endometrium; gene expression; gene induction /repression; gene mutation; genetic library; neoplasm /cancer genetics; neoplastic transformation; polymerase chain reaction; technology /technique development; tissue /cell culture; uterus neoplasms

DESCRIPTION (provided by applicant): Endometrial cancer is the most abundant cancer of the genital tract in American women and ranks fourth in overall incidence among cancers in women. Therefore, discoveries that contribute significantly to our understanding of this disease and hat provide new methods with which novel therapies may be discovered, would have a high impact on cancer in women. In this application we propose a pilot study to culture normal endometrial epithelial cells, which have very limited longevity in culture, and derive cultures from them that have a greatly increased longevity and capacity for proliferation in vitro. In the past we did with transfers of SV40 large T antigen but the cells that result are notably abnormal, behaving like severely hyperplastic to early malignant endometrial cells. Furthermore, we used an agent that appears to have no role in endometrial cancer development. In this pilot study we seek to develop propagable human endometrial epithelial cells that have been genetically modified in a manner that better approximates changes occurring during endometrial carcinogenesis. We seek to develop cells with an enhanced capacity for proliferation. To accomplish this we propose the following three specific aims 1) to determine the best genetic transfer agents and techniques applicable to primary cultures of normal epithelial endometrial cells. 2) to test the best way to impair PTEN function in these cells, recreating a recognized early event in endometrial carcinogenesis, and determine whether this change results in cells with increased capacity for cell proliferation. 3) to discover further mutations specifically relevant to development of endometrial cancer using various genetic libraries as "virtual mutagens." These studies will assess increased capacity for cell proliferation and will employ novel selection protocols that will mimic the selection pressures exerted by basement membrane and stromal cells in the uterine tissue. By identifying the alterations in cell physiology produced by this process, we hope to clarify the mechanisms that allow carcinogenic endometrial epithelial cells to escape the homeostatic controls on cell proliferation that normally operate in within endometrial tissue.

描述(申请人提供)：子宫内膜癌是美国女性生殖道中发病率最高的癌症，在女性癌症总发病率中排名第四。因此，有助于我们了解这种疾病的重大发现，以及为发现新疗法提供新方法的发现，将对女性癌症产生很大影响。在这项应用中，我们提出了一项初步研究，培养正常的子宫内膜上皮细胞，这些细胞在培养中的寿命非常有限，并从中衍生出极大地延长了寿命和体外增殖能力的培养。在过去，我们进行了SV40大T抗原的转移，但结果细胞明显异常，表现为严重增生到早期恶性子宫内膜细胞。此外，我们使用了一种似乎在子宫内膜癌发生中没有作用的药物。在这项先导性研究中，我们试图开发可繁殖的人类子宫内膜上皮细胞，这些细胞已经以一种更接近于子宫内膜癌变过程中发生的变化的方式进行了基因改造。我们寻求开发具有增强增殖能力的细胞。为此，我们提出了以下三个具体目标：1)确定适用于原代培养正常子宫内膜细胞的最佳基因转移剂和技术。2)测试在这些细胞中损害PTEN功能的最佳方法，重建已知的子宫内膜癌变的早期事件，并确定这种变化是否会导致细胞增殖能力增强。3)利用不同的基因文库作为“虚拟诱变剂”，进一步发现与子宫内膜癌发生有关的突变。这些研究将评估细胞增殖能力的提高，并将采用新的选择方案，模拟子宫组织中基底膜和基质细胞施加的选择压力。通过确定这一过程引起的细胞生理变化，我们希望阐明致癌子宫内膜上皮细胞逃脱细胞增殖的动态平衡控制的机制，这种控制通常在子宫内膜组织内运行。