PHENOL RED IN TISSUE-CULTURE MEDIA IS A WEAK ESTROGEN - IMPLICATIONS CONCERNING THE STUDY OF ESTROGEN-RESPONSIVE CELLS IN CULTURE
PHENOL RED IN TISSUE-CULTURE MEDIA IS A WEAK ESTROGEN - IMPLICATIONS CONCERNING THE STUDY OF ESTROGEN-RESPONSIVE CELLS IN CULTURE
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DOI：
10.1073/pnas.83.8.2496
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发表时间：
1986-04-01
影响因子：
11.1
通讯作者：
KATZENELLENBOGEN, BS
中科院分区：
文献类型：
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作者：
BERTHOIS, Y;KATZENELLENBOGEN, JA;KATZENELLENBOGEN, BS
尽管从血清中去除激素和开发无血清培养基来研究激素反应细胞已经引起了很大的关注，但很少考虑到培养基组分本身具有激素活性的可能性。我们发现，酚红在结构上与一些非甾体雌激素相似，在组织培养介质中被广泛用作pH指示剂，在组织培养介质中发现的浓度(15-45µm)具有显著的雌激素样活性。酚红与人乳腺癌细胞的雌激素受体结合的亲和力是雌二醇的0.001%(Kd=2。10-5M)。对雌激素受体阳性的乳腺癌细胞MCF-7的增殖有促进作用，且呈剂量依赖性，但对雌激素受体阴性的乳腺癌细胞MDA-MB-231的生长无影响。在组织培养液中存在的浓度下，酚红可引起部分雌激素刺激，使细胞数量增加到无酚红培养细胞的200%，孕激素受体含量增加到300%，从而降低外源雌激素刺激反应的程度。只有当细胞在酚红的存在下生长时，抗雌激素他莫昔芬和羟基他莫昔芬才能将细胞的增殖抑制在对照水平以下；在没有酚红的情况下，抗雌激素不会抑制细胞的生长。在任何利用雌激素反应细胞进行培养的研究中，都应该考虑酚红的雌激素活性。
Although much attention has been paid to the removal of hormones from sera and to the development of serum-free media for studies on hormone-responsive cells in culture, little consideration has been given to the possibility that the media components themselves may have hormonal activity. We have found that phenol red, which bears a structural resemblance to some nonsteroidal estrogens and which is used ubiquitously as a pH indicator in tissue culture media, has significant estrogenic activity at the concentrations (15-45 .mu.M) at which it is found in tissue culture media. Phenol red binds to the estrogen receptor of MCF-7 human breast cancer cells with an affinity 0.001% that of estradiol (Kd = 2 .times. 10-5 M). It stimulates the proliferation of estrogen receptor-positive MCF-7 breast cancer cells in a dose-dependent manner but has no effect on the growth of estrogen receptor-negative MDA-MB-231 breast cancer cells. At the concentrations present in tissue culture media, phenol red causes partial estrogenic stimulation, increasing cell number to 200% and progesterone receptor content to 300% of that found for cells grown in phenol red-free media, thereby reducing the degree to which exogenous estrogen is able to stimulate responses. The anti-estrogens tamoxifen and hydroxytamoxifen inhibit cell proliferation below the control level only when cells are grown in the presence of phenol red; in the absence of phenol red, the antiestrogens do not suppress growth. The estrogenic activity of phenol red should be considered in any studies that utilize estrogen-responsive cells in culture.