Human melanoblasts in culture: Expression of BRN2 and synergistic regulation by fibroblast growth factor-2, stem cell factor, and endothelin-3
Human melanoblasts in culture: Expression of BRN2 and synergistic regulation by fibroblast growth factor-2, stem cell factor, and endothelin-3
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DOI：
10.1046/j.1523-1747.2003.12562.x
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发表时间：
2003-11-01
影响因子：
6.5
通讯作者：
Sturm, RA
中科院分区：
文献类型：
--
作者：
Cook, AL;Donatien, PD;Sturm, RA
BRN 2转录因子（POU 3F 2，N-Oct-3）与黑色素细胞谱系的发育和黑色素瘤有关。使用低钙培养基补充干细胞因子，成纤维细胞生长因子-2，内皮素-3和霍乱毒素，我们已经建立和部分特征的人黑素细胞前体细胞，这是无色素，含有不成熟的黑素体和缺乏L-二羟基苯丙氨酸反应。与黑素细胞相比，成黑素细胞培养物表达高水平的BRN 2，当在含有佛波酯但缺乏内皮素-3、干细胞因子和成纤维细胞生长因子-2的培养基中培养时，BRN 2降低至与黑素细胞相似的水平。BRN 2的这种减少伴随着阳性L-二羟基苯丙氨酸反应和黑素体成熟的诱导，这与发育过程中观察到的成黑素细胞分化一致。在补充有干细胞因子、成纤维细胞生长因子-2和内皮素-3的低钙培养基中培养原代黑素细胞引起BRN 2蛋白水平增加，伴随着成黑素细胞样形态的变化。这些生长因子中的任何两个之间的协同作用是BRN 2蛋白诱导所必需的，而所有三个因子都是改变黑素细胞形态和最大化BRN 2蛋白表达所必需的。这些发现暗示BRN 2作为成黑素细胞的早期标志物，可能有助于黑素细胞基因控制的层次结构。
The BRN2 transcription factor (POU3F2, N-Oct-3) has been implicated in development of the melanocytic lineage and in melanoma. Using a low calcium medium supplemented with stem cell factor, fibroblast growth factor-2, endothelin-3 and cholera toxin, we have established and partially characterised human melanocyte precursor cells, which are unpigmented, contain immature melanosomes and lack L-dihydroxyphenylalanine reactivity. Melanoblast cultures expressed high levels of BRN2 compared to melanocytes, which decreased to a level similar to that of melanocytes when cultured in medium that contained phorbol ester but lacked endothelin-3, stem cell factor and fibroblast growth factor-2. This decrease in BRN2 accompanied a positive L-dihydroxyphenylalanine reaction and induction of melanosome maturation consistent with melanoblast differentiation seen during development. Culture of primary melanocytes in low calcium medium supplemented with stem cell factor, fibroblast growth factor-2 and endothelin-3 caused an increase in BRN2 protein levels with a concomitant change to a melanoblast-like morphology. Synergism between any two of these growth factors was required for BRN2 protein induction, whereas all three factors were required to alter melanocyte morphology and for maximal BRN2 protein expression. These finding implicate BRN2 as an early marker of melanoblasts that may contribute to the hierarchy of melanocytic gene control.