TRANSFORMATION OF HUMAN BREAST EPITHELIAL CELLS IN VITRO

人乳腺上皮细胞的一致性恶性转化 细胞或其他人类细胞，在体外通过化学或物理方法 致癌物质的研究至今尚未完成。 在可能的 失败的原因是1）使用不合适的目标 用于转化研究的细胞，即分化细胞 而不是干细胞; 2）使用不适当的实验室 动物作为转化细胞的宿主。 的主要目标 这份拨款提案是为了检验这两个问题的有效性， 可能性 最近我们观察到一个商场的存在 人肾上皮细胞亚群，含干细胞 特征，即，延长使用寿命，并能够接触- X射线致死照射人成纤维细胞不敏感生长 细胞垫 这和我们的初步结果表明， 可以使用细胞来富集人类乳房更新能力， MAT或适当的细胞培养基。 因此，我们建议 用化学和物理致癌物转化这些细胞， 通过使用1）我们的X射线， 致死性辐照的人成纤维细胞垫，另外， 已经显示出允许各种类型的人类癌的生长， 起源而非正常细胞（除了接触不敏感的 将对推定的转化细胞进行生物学表征， 从生物化学上证明它们确实是肿瘤 转化. 该提案的第二个主要目标是发展 能够接受和维持转化的新体内模型 乳腺上皮细胞 移植成功率和 原发性乳腺癌（大鼠和人）的生长速率， 推定的体外转化的人乳腺上皮细胞将 在3种免疫缺陷小鼠品系中进行检查，即， 常规无胸腺裸鼠（T淋巴细胞缺陷），beige-XID 无胸腺裸鼠（T淋巴细胞缺陷和LAK淋巴细胞）。 免疫缺陷小鼠的内分泌系统将被改变 提供一个富含人类乳房营养素的荷尔蒙环境， 荷尔蒙 在这项研究计划中，我们认为， 解决和研究复杂的两个非常关键的组成部分， 人乳腺的离体肿瘤转化过程 上皮细胞，过去未接受 足够的实验注意力。

Consistent malignant transformation of human breast epithelial cells, or other human cells, in vitro by chemical or physical carcinogens has not been accomplished to date. Among the possible reasons for the failure are 1) the use of inappropriate target cells for transformation studies, i.e. differentiating cells instead of stem cells; and 2) the use of inappropriate laboratory animals as hosts for transformed cells. The major objective of this grant proposal is to examine the validity of these two possibilities. Recently we have observed the existence of a mall subpopulation of human kidney epithelial cells, with stem cell characteristics, i.e., extended life span and capable of contact- insensitive growth on a x-ray lethally irradiated human fibroblast cell mat. This, and our preliminary results, indicate that it is possible to enrich for human breast renewal capacity using the cell mat or an appropriate cell culture medium. Therefore we propose to transform these cells with chemical and physical carcinogens and to select neoplastic transformed cells by using 1) our x-ray lethally irradiated human fibroblast cell mat which, additionally, has been shown to allow the growth of human carcinomas of various origin but not normal cells (except the contact-insensitive putative transformed cells will be characterized biologically and biochemically to demonstrate that they are truly neoplastic transformed. The second major goal of this proposal is to develop new in vivo models capable of accepting and maintaining transformed mammary epithelial cells. The transplantation success rate and growth rate of primary mammary carcinoma (rat and human) and putative in vitro transformed human breast epithelial cells will be examined in 3 strains of immune deficient mice, i.e., conventional athymic nude mice (T-lymphocyte deficient), beige-XID athymic nude mice (T-lymphocyte deficient and LAK lymphocytes). The endocrine system of the immune deficient mice will be altered to provide a hormonal milieu that is rich in human mammotrophic hormones. In this research proposal, we feel that we are addressing and examining two very crucial components of the complex process of ex vivo neoplastic transformation of human breast epithelium, components that have not in the past received sufficient experimental attention.