羽扇豆醇可能通过调控Notch信号通路抑制肺癌A549细胞恶性进展

Inhibition of lupinol on the malignant progression of lung cancer A549 cells through regulating Notch signaling pathway

目的探讨羽扇豆醇通过调控Notch信号通路对肺癌细胞增殖、凋亡、迁移及侵袭的机制。方法体外培养肺癌细胞A549,用浓度为0、15、30、60 mg/L的羽扇豆醇处理细胞,其中0 mg/L记对照组,其余记为羽扇豆醇各剂量组;采用浓度为60 mg/L羽扇豆醇和浓度为10μmol/L Notch通路抑制剂DAPT处理细胞,记为羽扇豆醇+DAPT组。通过四甲基偶氮唑盐(MTT)检测细胞增殖变化;流式细胞术检测细胞凋亡;Transwell检测细胞迁移和侵袭;Western blot检测细胞增殖核抗原(PCNA)、B淋巴细胞瘤-2(Bcl-2)、神经钙粘素(N-cadherin)、上皮钙粘素(E-cadherin)、Notch-1、发状分裂相关增强子(Hes-1)蛋白表达。结果15、30、60 mg/L羽扇豆醇组能明显抑制肺癌细胞的活力、迁移和侵袭细胞数目,提高凋亡率,降低PCNA、N-cadherin、Bcl-2、Hes-1、Notch-1表达,上调E-cadherin表达(P<0.05)。与羽扇豆醇组相比,羽扇豆醇+DAPT组明显降低细胞活力、迁移细胞数目、侵袭细胞数目,增加细胞凋亡率,降低PCNA、Bcl-2、Hes-1、Notch-1、N-cadherin蛋白表达,增加E-cadherin蛋白表达(P<0.05)。结论羽扇豆醇可能通过Notch信号通路阻碍肺癌细胞侵袭、迁移和增殖,诱导凋亡。

Objective To explore the mechanism of lupinol on the proliferation,apoptosis,migration and invasion of lung cancer cells through regulating Notch signaling pathway.Methods Lung cancer cells A549 were cultured in vitro,and the cells were treated with lupeol at concentrations of 0,15,30,and 60 mg/L,with 0 mg/L being the control group and the rese being the lupeol dosage groups,the cells were treated with lupinol at the concentration of 60 mg/L and Notch pathway inhibitor DAPT at the concentration of 10μmol/L,which was recorded as the lupeol+DAPT group.Cell proliferation changes were detected by MTT;cell apoptosis was detected by flow cytometry;cell migration and invasion were detected by Transwell;protein expression of PCNA,Bcl-2,N-cadherin,E-cadherin,Notch-1 and Hes 1 were detected by Western blot.Results 15,30,60 mg/L lupeol group can significantly inhibit the cell viability,the number of migration cells and invading cells of lung cancer cells,significantly increase the rate of cell apoptosis,and reduce PCNA,N-cadherin,Bcl-2,Hes-1 and Notch-1 expression,increase E-cadherin expression(P<0.05).Compared with the lupeol group,the lupeol+DAPT group significantly reduced cell viability,the number of migrating cells and invaded cells,increased apoptosis rate,decreased PCNA,Bcl-2,Hes-1,Notch-1 and N-cadherin protein expression,and increased E-cadherin protein expression(P<0.05).Conclusion Lupinol may inhibit the invasion,migration and proliferation of lung cancer cells through Notch signaling pathway,and induce apoptosis.