摘要
目的 探究安罗替尼通过调控核因子κB(NF-κB)信号通路对脑胶质瘤细胞恶性表型的影响.方法 体外培养人脑胶质瘤T98G细胞,以5-氟尿嘧啶为阳性对照药物,考察不同浓度(5、10、20 μmol/L)安罗替尼对该细胞增殖、黏附、迁移、侵袭能力和上皮间质转化(EMT)相关蛋白[上皮钙黏着蛋白(E-cadherin)、神经钙黏着蛋白(N-cadherin)、波形蛋白(vimentin)、纤维连接蛋白(FN)]表达的影响,并通过加入NF-κB信号通路抑制剂(BAY 11-7082)和激活剂(prostratin)来验证安罗替尼上述作用的可能机制.结果 5、10、20 μmol/L的安罗替尼均可显著降低细胞的增殖活力(5 μmol/L安罗替尼组除外)和迁移率,显著减少黏附细胞数和侵袭细胞数,显著上调E-cadherin蛋白的表达并下调N-cadherin、vimentin、FN蛋白的表达(P＜0.05),且20 μmol/L安罗替尼的作用与阳性对照药物相当(P＞0.05);与10 μmol/L安罗替尼比较,通路抑制剂可使细胞增殖、黏附、迁移、侵袭能力以及N-cadherin、vimentin、FN、磷酸化NF-κB p65蛋白的表达显著降低,E-cadherin蛋白的表达显著上调(P＜0.05),而通路激活剂则可使上述指标显著逆转(P＜0.05).结论 安罗替尼可抑制人脑胶质瘤T98G细胞的增殖、黏附、迁移和侵袭,上述作用可能与通过抑制NF-κB信号通路进而抑制细胞EMT样进程有关.
Abstract
OBJECTIVE To investigate the effects of anlotinib on the malignant phenotype of glioma cells by regulating the nuclear factor-κB(NF-κB)signaling pathway.METHODS Human glioma T98G cells were cultured in vitro,and 5-fluorouracil was used as positive control to investigate the effects of different concentrations of anlotinib(5,10,20 μmol/L)on the ability of proliferation,adhesion,migration and invasion,the expressions of epithelial-mesenchymal transition(EMT)related proteins[E-cadherin,N-cadherin,vimentin and fibronectin(FN)].NF-κB signaling pathway inhibitor(BAY 11-7082)and activator(prostratin)were additionally used to verify the possible mechanism of the above effects of anlotinib.RESULTS Anlotinib with 5,10,20 μmol/L could significantly decrease the activity of cell proliferation(except for 5 μmol/L anlotinib group),migration rate,and the number of adherent cells and invasive cells,could significantly up-regulate the expression of E-cadherin protein while down-regulate the expressions of N-cadherin,vimentin and FN protein(P＜0.05);the effect of 20 μmol/L anlotinib was similar to that of positive control(P＞0.05).Compared with 10 μmol/L anlotinib,pathway inhibitor could significantly decrease the ability of proliferation,adhesion,migration and invasion,and the expressions of N-cadherin,vimentin,FN and phosphorylated NF-κB p65 protein,while could significantly up-regulate the expression of E-cadherin protein(P＜0.05);above indexes were reversed significantly by pathway activator(P＜0.05).CONCLUSIONS Anlotinib may inhibit the proliferation,adhesion,migration and invasion of human glioma T98G cells,which may be associated with the inhibition of the NF-κB signaling pathway,thus inhibiting cell EMT-like processes.
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维普
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