Richards J, Imagawa W, Balakrishnan A, Edery M, Nandi S
Cancer Research Laboratory, University of California, Berkeley 94720.
Endocrinology. 1988 Sep;123(3):1335-40. doi: 10.1210/endo-123-3-1335.
Normal and neoplastic mammary cells from both human and rodent sources grow in culture in response to a number of hormones and growth factors. However, with the exception of a few human tumor lines, a consistent growth-promoting effect of estrogens on mammary cells has not been observed. Mammary cells can be shown to respond to other hormones and factors, such as PRL, hydrocortisone, and epidermal growth factor. Recent observations suggest that the pH indicator dye phenol red, found in most media, may be masking any exogenous estrogenic effects by acting as a weak estrogen. To test this possibility, we reexamined the effects of estradiol (E2) and the antiestrogen keoxifene on the growth of normal human, mouse, and rat mammary cells in the absence of phenol red. Primary cultures of these mammary cells were grown within a rat tail collagen gel matrix in a serum-free medium made up of Ham's F-12 and Dulbecco's Modified Eagles' medium (1:1) with and without phenol red. The medium was supplemented with various hormones and growth factors. These supplements were selected for each cell type to produce a variety of conditions from nongrowing to rapidly growing. The effects of E2 (10(-10)-10(-8) M, keoxifene (10(-9)-10(-6) M), and phenol red on growth under these various conditions were examined. Phenol red had no effect on growth, and its absence did not restore a response to E2. Keoxifene, in the presence or absence of E2, also had no effect on growth. Although E2 had no effect on growth, it was able to induce a 150% increase in the progesterone receptor levels in normal mouse mammary cells in culture, indicating that the cells retain their capacity to respond to E2. This work supports the idea that the effect of estrogens on growth in vivo may be mediated through some other factor(s).
来自人类和啮齿动物的正常乳腺细胞及肿瘤性乳腺细胞在培养中会对多种激素和生长因子产生反应。然而，除了少数人类肿瘤细胞系外，尚未观察到雌激素对乳腺细胞具有一致的促生长作用。乳腺细胞可被证明对其他激素和因子有反应，如催乳素、氢化可的松和表皮生长因子。最近的观察表明，大多数培养基中含有的pH指示剂染料酚红可能作为一种弱雌激素掩盖了任何外源性雌激素的作用。为了验证这种可能性，我们在不含酚红的情况下重新研究了雌二醇（E2）和抗雌激素药物凯西芬对正常人、小鼠和大鼠乳腺细胞生长的影响。这些乳腺细胞的原代培养物在大鼠尾胶原凝胶基质中，于由Ham's F - 12和杜尔贝科改良伊格尔培养基（1:1）组成的无血清培养基中生长，该培养基含或不含酚红。培养基中添加了各种激素和生长因子。针对每种细胞类型选择这些添加物，以产生从非生长到快速生长的各种条件。研究了E2（10^(-10) - 10^(-8) M）、凯西芬（10^(-9) - 10^(-6) M）和酚红在这些不同条件下对生长的影响。酚红对生长无影响，且去除酚红也未恢复对E2的反应。无论有无E2存在，凯西芬对生长也无影响。尽管E2对生长无影响，但它能够使培养的正常小鼠乳腺细胞中的孕激素受体水平增加150%，这表明细胞仍保留对E2作出反应的能力。这项工作支持了雌激素对体内生长的影响可能通过某些其他因素介导的观点。
