Departments of Pediatrics, Neurology, and Neurobiology & Anatomy, University of Rochester Medical Center, 601 Elmwood Avenue, Box 777, Rochester, NY 14642, USA.
Exp Cell Res. 2013 Mar 10;319(5):660-9. doi: 10.1016/j.yexcr.2012.12.027. Epub 2013 Jan 16.
Peripheral neuroblastic tumors exist as a heterogeneous mixture of neuroblastic (N-type) cells and Schwannian stromal (S-type) cells. These stromal cells not only represent a differentiated and less aggressive fraction of the tumor, but also have properties that can influence the further differentiation of nearby malignant cells. In vitro neuroblastoma cultures exhibit similar heterogeneity with N-type and S-type cells representing the neuroblastic and stromal portions of the tumor, respectively, in behavior, morphology, and molecular expression patterns. In this study, we deplete kinase D-interacting substrate of 220kD (Kidins220) with an shRNA construct and thereby cause morphologic transition of the human SH-SY5Y neuroblastoma cell line from N-type to S-type. The resulting cells have similar morphology and expression profile to SH-EP1 cells, a native S-type cell line from the same parent cell line, and to SH-SY5Y cells treated with BrdU, a treatment that induces S-type morphology. Specifically, both Kidins220-deficient SH-SY5Y cells and native SH-EP1 cells demonstrate down-regulation of the genes DCX and STMN2, markers for the neuronal lineage. We further show that Kidins220, DCX and STMN2 are co-down-regulated in cells of S-type morphology generated by methods other than Kidins220 depletion. Finally, we report that the association of low Kidins220 expression with S-type morphology and low DCX and STMN2 expression is demonstrated in spontaneously occurring human peripheral neuroblastic tumors. We propose that Kidins220 is critical in N- to S-type transition of neural crest tumor cells.
外周神经母细胞瘤是一种神经母细胞（N 型）和许旺氏基质（S 型）细胞组成的异质性混合物。这些基质细胞不仅代表肿瘤中分化程度较低且侵袭性较弱的部分，而且具有影响附近恶性细胞进一步分化的特性。体外神经母细胞瘤培养物表现出相似的异质性，N 型和 S 型细胞分别代表肿瘤的神经母细胞和基质部分，在行为、形态和分子表达模式上具有相似性。在本研究中，我们使用 shRNA 构建体耗尽激酶 D 相互作用的底物 220kD（Kidins220），从而导致人 SH-SY5Y 神经母细胞瘤细胞系从 N 型向 S 型形态发生转变。由此产生的细胞具有与 SH-EP1 细胞相似的形态和表达谱，SH-EP1 细胞是同源于同一亲本细胞系的 S 型细胞系，与用 BrdU 处理的 SH-SY5Y 细胞相似，BrdU 处理诱导 S 型形态。具体而言，Kidins220 缺陷的 SH-SY5Y 细胞和天然 SH-EP1 细胞均表现出神经元谱系标志物 DCX 和 STMN2 基因的下调。我们进一步表明，除了 Kidins220 耗竭之外，通过其他方法产生的 S 型形态的细胞中，Kidins220、DCX 和 STMN2 均共同下调。最后，我们报告说，在自发发生的人类外周神经母细胞瘤中，低水平的 Kidins220 表达与 S 型形态和低水平的 DCX 和 STMN2 表达相关。我们提出 Kidins220 在外周神经母细胞瘤细胞从 N 型向 S 型转变中起关键作用。
