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Study of transforming growth factor β in melanocyte proliferation and differentiation of mouse neural crest cells via stem cell factor/KIT signaling

通过干细胞因子/KIT信号研究转化生长因子β在小鼠神经嵴细胞黑素细胞增殖和分化中的作用

基本信息

批准号：
14570828
负责人：
KAWAKAMI Takahiro
金额：
$ 2.5万
依托单位：
St.Marianna University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570828/
关键词：
TGF β neural crest cells melanocyte mouse NCC malignant melanoma differentiation SCF KIT proliferation TGFβ

项目摘要

SCF is essential to the migration and differentiation of melanocytes during embryogenesis based on the observation that mutations in either the SCF gene, or its ligand, KIT, result in defects in coat pigmentation in mice. TGF β1 has been implicated in the regulation of both cellular proliferation and differentiation. NCC-melb4, an immortal cloned cell line, was cloned from a mouse NCC. NCC-melb4 cells provide model to study the specific stage of differentiation and proliferation of melanocytes. They also express KIT as a melanoblast marker. Using the NCC-melb4 cell line, we investigated the effect of TGF β1 on the differentiation and proliferation of immature melanocyte precursors. Immunohistochemically, NCC-melb4 cells showed TGF β1 expression. The anti-TGF β1 antibody inhibited the cell growth, and downregulated the KIT protein and mRNA expression. To further investigate the activation of autocrine TGF β1, NCC-melb4 cells were incubated on-exogenous TGF β1 culture medium. KIT protein decreased with anti-TGF β1 antibody concentration in a dose-dependent fashion. We concluded that in NCC-melb4 cells, TGF β1 promotes melanocyte precursor proliferation in autocrine and/or paracrine regulation. We further investigated the influence of TGF β1 in vitro using a NCC primary culture system from wild-type mice. Anti-TGF β1 antibody decreased the number of KIT positive NCC. I addition, the anti-TGF β1 antibody supplied within the wild-type neural crest explants abolished the growth of NCC. These results indicate that TGF β1 affect melanocyte precursor proliferation and differentiation in the presence of SCF/KIT in an autocrine/paracrine manner.

SCF在胚胎发生过程中对黑素细胞的迁移和分化至关重要，这是基于SCF基因或其配体KIT突变导致小鼠皮毛色素沉着缺陷的观察结果。TGF β1参与细胞增殖和分化的调节。NCC-melb 4是从小鼠NCC克隆的永生克隆细胞系。NCC-melb 4细胞为研究黑素细胞分化和增殖的特定阶段提供了模型。它们还表达KIT作为成黑素细胞标记物。我们使用NCC-melb 4细胞系，研究了TGF β1对未成熟黑素细胞前体细胞分化和增殖的影响。免疫组化显示NCC-melb 4细胞表达TGF β1。抗TGF β1抗体抑制细胞生长，下调KIT蛋白和mRNA表达。为了进一步研究自分泌TGF β1的激活，将NCC-melb 4细胞在外源性TGF β1培养基上孵育。KIT蛋白随着抗TGF β1抗体浓度的增加呈剂量依赖性下降。我们的结论是，在NCC-melb 4细胞中，TGF β1通过自分泌和/或旁分泌调节促进黑素细胞前体增殖。我们使用野生型小鼠的NCC原代培养系统进一步研究了TGF β1的体外影响。抗TGF β1抗体可减少KIT阳性NCC的数量。此外，在野生型神经嵴外植体中提供的抗TGF β1抗体消除了NCC的生长。这些结果表明TGF β1在SCF/KIT存在下以自分泌/旁分泌方式影响黑素细胞前体细胞的增殖和分化。