Differential responsiveness of human breast cancer cell lines MCF-7 and T47D to growth factors and 17 beta-estradiol.
Differential responsiveness of human breast cancer cell lines MCF-7 and T47D to growth factors and 17 beta-estradiol.
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发表时间：
1988-07
期刊：
影响因子：
11.2
通讯作者：
Kenneth P. Karey;D. Sirbasku
中科院分区：
文献类型：
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作者：
Kenneth P. Karey;D. Sirbasku
一种完全无血清的测定方法已被用来比较多肽生长因子和雌激素与MCF-7和T47 D人乳腺癌细胞培养物中的促有丝分裂活性。将细胞系保持在Ham’s F12和Dulbecco’s改良Eagle’s培养基（1：1）中的存活的、缓慢分裂的条件下，所述培养基补充有碳酸氢钠（2.2 g/l）、15 mM 4-（2-羟乙基）-1-哌嗪乙烷磺酸、人转铁蛋白（10微克/ml）和牛血清白蛋白（200微克/ml）（指定为Tf/BSA）。该培养基允许通过多轮细胞分裂测定促有丝分裂活性，并允许比较功能家族内生长因子以及不同促有丝分裂剂的生物学效价。胰岛素样生长因子I（IGF-I）是研究的最有效的促有丝分裂剂，MCF-7和T47 D细胞的ED 50值分别为160 pg/ml和1.7 ng/ml。胰岛素样生长因子II和胰岛素活性较低，MCF-7细胞的ED 50值分别为0.55和1.2 ng/ml，T47 D细胞系的ED 50值分别为4.3和10 ng/ml。具有表皮生长因子样功能特性的促分裂原的ED 50值为35 pg/ml至2.5 ng/ml，而转化生长因子β型和血小板衍生生长因子没有可检测的刺激作用。碱性成纤维细胞生长因子对MCF-7和T47 D细胞的ED 50值分别为0.42 ng/ml和3.7 ng/ml，而酸性成纤维细胞生长因子几乎无活性。在不含酚红的Tf/BSA中，17 β-雌二醇在8天内导致MCF-7细胞数量比对照增加60%，而对T47 D细胞系的生长没有影响。从MCF-7条件Tf/BSA培养基中，IGF-I通过生物活性、通过放射免疫测定（约等于2 pg/ml）和通过在解离条件下估计分子量（8，000）来鉴定。IGF-I的浓度不受17 β-雌二醇治疗的影响。这些数据表明，诱导酸稳定，低分子量自分泌生长因子涉及更多的调节比单独雌激素定义。17 β-雌二醇在Tf/BSA中的最小作用开辟了几种可能性，包括其他血清传播的激素、生长因子和调节剂在自分泌生长因子诱导中的假定作用。
A completely serum-free assay method has been used to compare the mitogenic activities of polypeptide growth factors and estrogens with MCF-7 and T47D human breast cancer cells in culture. The lines were maintained in a viable, slowly dividing condition in Ham's F12 and Dulbecco's modified Eagle's medium (1:1) supplemented with sodium bicarbonate (2.2 g/liter), 15 mM 4-(2-hydroxyethyl)-1-piperazineethane-sulfonic acid, human transferrin (10 micrograms/ml), and bovine serum albumin (200 micrograms/ml) (designated Tf/BSA). This medium allowed the assay of mitogenic activities as measured by multiple rounds of cell division and permitted comparisons of the biological potencies of growth factors within functional families as well as of dissimilar mitogens. Insulin-like growth factor I (IGF-I) was the most potent mitogen studied, showing ED50 values of 160 pg/ml and 1.7 ng/ml with the MCF-7 and T47D cells, respectively. Insulin-like growth factor II and insulin were less active, with ED50 values of 0.55 and 1.2 ng/ml with MCF-7 cells and 4.3 and 10 ng/ml with the T47D cell line, respectively. Mitogens sharing epidermal growth factor-like functional properties had ED50 values from 35 pg/ml to 2.5 ng/ml, while transforming growth factor type beta and platelet-derived growth factor had no detectable stimulatory effects. Basic fibroblast growth factor had ED50 values of 0.42 ng/ml and 3.7 ng/ml for the MCF-7 and T47D cells, respectively, while acidic fibroblast growth factor was nearly inactive. In phenol red-free Tf/BSA, 17 beta-estradiol caused a 60% increase in MCF-7 cell numbers over controls in 8 days while having no effect on growth of the T47D cell line. From MCF-7 conditioned Tf/BSA medium, IGF-I was identified by biological activity, by radioimmunoassay (approximately equal to 2 pg/ml) and by estimation of molecular weight (8,000) under dissociating conditions. The concentration of IGF-I was not affected by 17 beta-estradiol treatment. The data indicate that induction of acid stable, low molecular weight autocrine growth factors involved more regulation than defined by estrogens alone. The minimal effects of 17 beta-estradiol in Tf/BSA opened several possibilities including the putative roles of other serum-borne hormones, growth factors and regulators in autocrine growth factor induction.