Collective cell migration plays an important role in embryonic development, wound healing, and cancer metastasis. We aimed to investigate the expression, role, and mechanism of Netrin-1 in collective cell migration using a3D culture model. An immunohistochemical study showed that certain cells invaded surrounding tissue by collective migration and that Netrin-1 expression in these cells was increased, especially at the invasive front. In the 3D culture model, collective cell migration was clearly observed, as leader cells were followed by cells migrating along a canal. N-cadherin-mediated cell junctions were observed in collective cell migration, and Netrin-1 expression was elevated in these cells. Netrin-1 did not affect the expression of N-cadherin in 2D-cultured cells; however, in 3D culture, the overexpression of Netrin-1 increased N-cadherin and promoted the collective migration of Huh7 cells, while the knockdown of Netrin-1 decreased N-cadherin and inhibited collective migration in SK-Hep-1 cells. Interestingly, N-cadherin knockdown in Huh7 cells significantly diminished Netrin-1-promoted collective cell migration, while the overexpression of N-cadherin restored collective migration in Netrin-1-knockdown SK-Hep1 cells. These results suggest that Netrin-1 enhances N-cadherin junctions to promote liver cancer cell collective migration in 3D cell culture and may subsequently increase liver cancer metastasis.

中文翻译：

---

Netrin-1在3D细胞培养模型中促进肝癌细胞的集体细胞迁移。

集体细胞迁移在胚胎发育，伤口愈合和癌症转移中起重要作用。我们旨在研究使用a3D培养模型的Netrin-1在集体细胞迁移中的表达，作用和机制。一项免疫组织化学研究表明，某些细胞通过集体迁移侵入周围组织，并且这些细胞中的Netrin-1表达增加，尤其是在侵袭前沿。在3D培养模型中，可以清楚地观察到集体细胞迁移，因为前导细胞随后是细胞沿管迁移。在集体细胞迁移中观察到N-钙黏着蛋白介导的细胞连接，并且Netrin-1表达在这些细胞中升高。Netrin-1不会影响2D培养细胞中N-钙粘蛋白的表达；但是，在3D文化中 Netrin-1的过表达增加了N-cadherin并促进了Huh7细胞的集体迁移，而敲除Netrin-1则降低了N-cadherin并抑制了SK-Hep-1细胞的集体迁移。有趣的是，Huh7细胞中的N-钙粘蛋白敲低显着减少了Netrin-1促进的集体细胞迁移，而N-钙粘蛋白的过表达恢复了Netrin-1敲除的SK-Hep1细胞中的集体迁移。这些结果表明，Netrin-1可增强N-钙粘蛋白连接，从而促进3D细胞培养中肝癌细胞的集体迁移，并可能随后增加肝癌的转移。在Huh7细胞中N-钙粘蛋白的敲低显着减少了Netrin-1促进的集体细胞迁移，而N-钙粘蛋白的过表达恢复了在Netrin-1敲除的SK-Hep1细胞中的集体迁移。这些结果表明，Netrin-1可增强N-钙粘蛋白连接，从而促进3D细胞培养中肝癌细胞的集体迁移，并可能随后增加肝癌的转移。在Huh7细胞中N-钙粘蛋白的敲低显着减少了Netrin-1促进的集体细胞迁移，而N-钙粘蛋白的过表达恢复了在Netrin-1敲除的SK-Hep1细胞中的集体迁移。这些结果表明，Netrin-1可增强N-钙粘蛋白连接，从而促进3D细胞培养中肝癌细胞的集体迁移，并可能随后增加肝癌的转移。