小檗胺通过OTUB1和MDM2-p53通路调控肺癌A549细胞的增殖、凋亡和侵袭转移

Berberamine Regulates the Proliferation,Apoptosis,Invasion and Metastasis of Lung Cancer A549 Cells Through the OTUB1 and MDM2-p53 Pathways

目的:探讨小檗胺对肺癌细胞A549的增殖、侵袭转移和凋亡的影响及可能的作用机制。方法:用MTT试验和EdU试剂盒检测小檗胺对A549细胞增殖的影响,划痕试验和Transwell试验检测小檗胺对A549细胞侵袭转移的影响,台盼蓝染色实验和Cell Death Detection Elisa Kit检测小檗胺对A549细胞凋亡的影响;Western blotting检测小檗胺对A549细胞中卵巢肿瘤相关蛋白酶B1(OTUB1)、鼠双微体2基因(MDM2)、p53及半胱氨酸蛋白酶-3(Caspase-3)表达的影响。结果:小檗胺以浓度依赖和时间依赖的形式抑制A549细胞增殖,在小檗胺处理A549细胞72 h时,小檗胺抑制细胞增殖的IC50为(22.7±2.3)μmol/L。与对照组比较,小檗胺能够明显诱导A549细胞凋亡(P<0.05)。划痕试验和Transwell试验结果表明小檗胺能够明显抑制A549细胞的侵袭转移(P<0.05)。Western blotting结果提示小檗胺能够明显下调OTUB1、MDM2的表达(P<0.05),上调p53和cleaved-Caspase-3/Caspase-3的表达(P<0.05)。结论:小檗胺能够抑制A549细胞增殖与侵袭转移,诱导细胞凋亡,这种作用可能是通过调控OTUB1和MDM2-p53信号通路来实现的。

Objective:To investigate the effect of berberamine on the proliferation,invasion,metastasis and apoptosis of lung cancer cell A549 and its possible mechanism.Methods:MTT test and EdU kit were used to detect the effect of berberamine on the proliferation of A549 cells.Scratch test and Transwell test were used to detect the effect of berberamine on the invasion and metastasis of A549 cells.Trypan blue staining and cell death detection ELISA kit were used to detect the effect of berberamine on the apoptosis of A549 cells.Western blotting was used to detect the effects of berberamine on the expression of OTUB1,MDM2,p53 and Caspase-3 in A549 cells.Results:Berberamine inhibited the proliferation of A549 cells in a concentration-dependent and time-dependent manner.When A549 cells were treated with berberamine for 72 h,the IC50 of berberamine inhibiting cell proliferation was(22.7±2.3)μmol/L.Compared with the control group,berberamine can significantly induce A549 cell apoptosis(P<0.05).The results of Scratch test and Transwell test showed that berberamine can significantly inhibit the invasion and metastasis of A549 cells(P<0.05).Western blotting results indicated that berberamine can significantly down-regulate the expression of OTUB1 and MDM2(P<0.05),and up-regulate the expression of p53 and cleaved-Caspase-3/Caspase-3(P<0.05).Conclusion:Berberamine can inhibit the proliferation,invasion and metastasis of A549 cells,and induce apoptosis.This effect may be achieved by regulating t he OTUB1 and MDM2-p53 signaling pathways.