Rogers S L, Cutts J L, Gegick P J, McGuire P G, Rosenberger C, Krisinski S
Department of Anatomy, University of New Mexico School of Medicine, Albuquerque 87131.
Exp Cell Res. 1994 Apr;211(2):252-62. doi: 10.1006/excr.1994.1085.
We reported previously (S. L. Rogers, P. J. Gegick, S. M. Alexander, and P. G. McGuire, Dev. Biol. 151, 191-203, 1992) that transforming growth factor-beta 1 (TGF beta 1) inhibited proliferation, up-regulated fibronectin synthesis, and suppressed melanogenesis in a population of quail neural crest cells in vitro. Here, we report that cell lines derived from the parent SK-N-SH neuroblastoma line (R. A. Ross, B. A. Spengler, and J. L. Biedler, J. Natl. Cancer Inst. 71, 741-747, 1983) respond differentially to TGF beta 1, and their responses provide further insights into the actions of this growth factor on neural crest subpopulations. The SH-EP cell line exhibits primarily nonneuronal traits and responded to TGF beta 1 with increased thymidine uptake after 6 days of culture, increased expression of fibronectin mRNA and protein, and decreased laminin synthesis. Many SH-EP cells also acquired a dramatically elongated morphology, reminiscent of Schwann cells in culture. Thymidine uptake by the neuronal SY5Y cell line was not substantially altered. Neither fibronectin mRNA nor protein was detectable in either TGF beta 1-treated or untreated cultures, although laminin synthesis was upregulated by the growth factor. In TGF beta 1-treated cultures of the intermediate SH-IN cell line, which has been reported to display both neuronal and nonneuronal characteristics, there was marked flattening of many cells, a steady decrease in thymidine uptake, and increased expression of both fibronectin and laminin. The observed responses of SH-IN cells mimic those observed in primary neural crest cultures and appear to represent similar differentiation toward a mesenchymal phenotype. These results substantiate the idea that closely related but diverging neural crest-derived cell types respond selectively to TGF beta 1 and demonstrate that these SK-N-SH-derived cell lines will be useful in experimental approaches that will allow us to infer mechanisms underlying regulation of neural crest differentiation.
我们之前报道过（S.L.罗杰斯、P.J.格吉克、S.M.亚历山大和P.G.麦圭尔，《发育生物学》151卷，第191 - 203页，1992年），转化生长因子β1（TGFβ1）在体外可抑制鹌鹑神经嵴细胞群体的增殖、上调纤连蛋白的合成并抑制黑色素生成。在此，我们报道源自亲代SK - N - SH神经母细胞瘤细胞系（R.A.罗斯、B.A.斯彭格勒和J.L.比德勒，《国家癌症研究所杂志》71卷，第741 - 747页，1983年）的细胞系对TGFβ1有不同反应，它们的反应为深入了解这种生长因子对神经嵴亚群的作用提供了更多线索。SH - EP细胞系主要表现出非神经元特征，培养6天后对TGFβ1的反应是胸苷摄取增加、纤连蛋白mRNA和蛋白表达增加以及层粘连蛋白合成减少。许多SH - EP细胞还呈现出显著拉长的形态，类似于培养中的施万细胞。神经元性的SY5Y细胞系的胸苷摄取没有明显改变。在TGFβ1处理或未处理的培养物中均未检测到纤连蛋白mRNA或蛋白，尽管层粘连蛋白合成被该生长因子上调。在中间型的SH - IN细胞系的TGFβ1处理培养物中，据报道该细胞系兼具神经元和非神经元特征，许多细胞出现明显扁平，胸苷摄取持续减少，纤连蛋白和层粘连蛋白的表达均增加。观察到的SH - IN细胞的反应类似于在原代神经嵴培养物中观察到的反应，似乎代表了向间充质表型的类似分化。这些结果证实了这样一种观点，即密切相关但不同的神经嵴衍生细胞类型对TGFβ1有选择性反应，并表明这些源自SK - N - SH的细胞系将有助于实验研究，使我们能够推断神经嵴分化调控的潜在机制。
