安罗替尼对脑胶质瘤T98G细胞迁移侵袭及上皮间质转化的抑制作用

目的探究安罗替尼对人脑胶质瘤细胞黏附、迁移、侵袭和上皮间质转化（EMT）的影响。方法体外培养人脑胶质瘤T98G细胞，分为对照组（不做干预）、实验组（2.5 μmol·L-1安罗替尼组、5 μmol·L-1安罗替尼组、10 μmol·L-1安罗替尼组）和阳性药物组（50 mg·L-1 5-氟尿嘧啶），干预24 h。用活细胞计数（CCK-8）、细胞黏附实验、划痕法、Transwell小室及蛋白免疫印迹（WB）法对细胞增殖活力、黏附、迁移、侵袭能力及EMT相关蛋白表达量进行分析。结果与对照组比较，5 μmol·L-1、10 μmol·L-1安罗替尼组和阳性药物组增殖活力显著降低（P<0.05），实验组和阳性药物组T98G细胞黏附、迁移、侵袭能力，纤维粘连蛋白（FN）、波形蛋白（Vimentin）、N-钙黏蛋白（N-cadherin）蛋白的表达水平呈现显著下调趋势（P<0.05），而E-钙黏蛋白（E-cadherin）蛋白的表达水平呈现明显上调趋势（P<0.05），且浓度越高变化越显著；与阳性药物组相比，2.5 μmol·L-1、5 μmol·L-1安罗替尼组细胞增殖活力、黏附、迁移、侵袭能力，N-cadherin、Vimentin及FN的蛋白水平上调（P<0.05），而E-cadherin蛋白水平则下调（P<0.05）；10 μmol·L-1安罗替尼组与阳性药物组相比无显著差异（P>0.05）。结论安罗替尼可能是通过抑制EMT进程进而抑制人脑胶质瘤T98G细胞的增殖、黏附、迁移、侵袭能力。

Objective To investigate the effects of anlotinib on the adhesion, migration, invasion and epithelial-mesenchymal transition(EMT) of human glioma cells. Methods Human glioma T98G cells were cultured in vitro and divided into control group(no intervention), experimental group(2.5 μmol·L-1 anlotinib group, 5 μmol·L-1 anlotinib group, 10 μmol·L-1 anlotinib group) and positive drug group(50 mg·L-1 5-fluorouracil) for 24 hours.Cell proliferation, adhesion, migration, invasion and EMT-related protein expression were analyzed by cell count(CCK-8), cell adhesion assay, scratch assay, Transwell and Western blotting(WB). Results Compared with the control group, the proliferative activity of 5 μmol·L-1, 10 μmol·L-1 and positive drug groups was significantly decreased(P<0.05), and the adhesion, migration and invasion ability of T98G cells in the experimental group and positive drug group were significantly decreased(P<0.05). The expression levels of fibrin(FN), Vimentin and N-cadherin were significantly down-regulated(P<0.05), while the expression levels of E-cadherin were significantly up-regulated(P<0.05). The higher the concentration, the more significant the change. Compared with positive drug group, the cell proliferation activity, adhesion, migration invasion ability, and the protein levels of N-cadherin, Vimentin and FN were up-regulated, while the protein level of E-cadherin was down-regulated in the 2.5 μmol·L-1 and 5 μmol·L-1 anlotinib groupsd(P<0.05). There was no significant difference between the 10 μmol·L-1 antirotinib group and the positive group(P>0.05). Conclusion Anlotinib may inhibit the proliferation, adhesion, migration and invasion ability of human glioma T98G cells by inhibiting the process of EMT.