Ding Xionghui, Xiang Li, Wang Ning, Zhao Zhanbo, Jin Xin, Sun Yanhui, Duan Wenjuan, Wang Shiqi, Jin Xianqing
Ministry of Education Key Laboratory of Child Development and Disorders, Yuzhong, Chongqing 400014, P.R. China.
Department of Neonatal Gastrointestinal Surgery, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, P.R. China.
Oncol Rep. 2014 Mar;31(3):1165-74. doi: 10.3892/or.2013.2963. Epub 2013 Dec 31.
Rearranged during transfection (RET) is widely expressed in neuroblastoma (NB) and partly contributes to high metastatic potential and survival of NB. The aim of the present study was to investigate whether vandetanib (a RET inhibitor) inhibits proliferation, migration and invasion of NB cells in vitro. The effects of vandetanib on the proliferation, apoptosis and cell cycle and on RET phosphorylation of SK-N-SH and SH-SY5Y cells were evaluated in vitro. The migration and invasion potential of vandetanib-treated NB cells were analyzed using Transwell cell migration and invasion assays, respectively. qPCR, western blotting and immunofluorescence were used to detect mRNA and protein levels in NB cells treated with vandetanib. Our data demonstrated that vandetanib inhibits the proliferation of SK-N-SH and SH-SY5Y cells and that this inhibition is mediated by the induction of G1 phase cell cycle arrest at lower concentrations and by apoptosis at higher concentrations. In the presence of vandetanib, the migration and invasion of two NB cell lines were markedly decreased compared with the control group (p<0.01). In addition, our data showed that the levels of C-X-C chemokine receptor type 4 (CXCR4) and matrix metalloproteinase 14 (MMP14) mRNA expression in NB cell lines treated with vandetanib were significantly lower than those in the cells that were treated with vehicle (p<0.01) and similar results were obtained for protein levels as determined by western blotting and immunofluorescence analysis. Vandetanib may inhibit the proliferation, migration and invasion of NB cells in vitro. The potential mechanisms for the inhibition of NB migration and invasion by vandetanib may partly be attributed to the ability of vandetanib to suppress the expression of CXCR4 and MMP14 in human NB cells.
转染期间重排（RET）在神经母细胞瘤（NB）中广泛表达，部分促成了NB的高转移潜能和生存能力。本研究的目的是调查凡德他尼（一种RET抑制剂）是否在体外抑制NB细胞的增殖、迁移和侵袭。在体外评估了凡德他尼对SK-N-SH和SH-SY5Y细胞的增殖、凋亡、细胞周期以及RET磷酸化的影响。分别使用Transwell细胞迁移和侵袭试验分析了经凡德他尼处理的NB细胞的迁移和侵袭潜能。采用qPCR、蛋白质印迹法和免疫荧光法检测经凡德他尼处理的NB细胞中的mRNA和蛋白质水平。我们的数据表明，凡德他尼抑制SK-N-SH和SH-SY5Y细胞的增殖，且这种抑制在较低浓度下由G1期细胞周期阻滞介导，在较高浓度下由凋亡介导。在凡德他尼存在的情况下，与对照组相比，两种NB细胞系的迁移和侵袭明显减少（p<0.01）。此外，我们的数据显示，经凡德他尼处理的NB细胞系中C-X-C趋化因子受体4（CXCR4）和基质金属蛋白酶14（MMP14）的mRNA表达水平显著低于用溶媒处理的细胞（p<0.01），蛋白质印迹法和免疫荧光分析测定的蛋白质水平也得到了类似结果。凡德他尼可能在体外抑制NB细胞的增殖、迁移和侵袭。凡德他尼抑制NB迁移和侵袭的潜在机制可能部分归因于凡德他尼抑制人NB细胞中CXCR4和MMP14表达的能力。
