番泻苷B抑制STAT3和ERK1/2磷酸化对A549细胞生长侵袭及裸鼠成瘤的影响

Effects of sennoside B on growth and invasion of A549 cells and tumorigenesis of nude mice by inhibiting the phosphorylation of STAT3 and ERK1/2

目的探讨番泻苷B对A549细胞生长、侵袭及裸鼠成瘤的影响及机制。方法采用0、5、10、20μM番泻苷B处理非小细胞肺癌A549细胞,将细胞随机分为4组进行后续实验,Brdu染色检测各组细胞增殖;Hoechst染色检测细胞凋亡;划痕实验检测细胞迁移;Transwell实验检测细胞侵袭;蛋白免疫印迹检测ki67、PCNA、cl-caspase-3、cl-caspase-9、VEGF、N-cadherin和E-cadherin蛋白表达水平,STAT3和ERK1/2的磷酸化情况;建立荷瘤小鼠模型,检测肿瘤重量,免疫组化检测Ki67和VEGF表达。结果与Control组相比较,各番泻苷B剂量组Brdu阳性细胞数量、侵袭细胞数明显减少(P<0.05),细胞凋亡率明显上升(P<0.05),细胞划痕愈合率降低(P<0.05),ki67、PCNA、VEGF、N-cadherin蛋白水平明显降低(P<0.05),cl-caspase-3、cl-caspase-9、E-cadherin蛋白水平明显升高(P<0.05),STAT3和ERK1/2的磷酸化水平均明显降低(P<0.05),降低荷瘤小鼠肿瘤重量与Ki67和VEGF表达水平(P<0.05)。结论番泻苷B抑制STAT3、ERK1/2磷酸化对A549细胞体内外生长有抑制作用。

Objective To investigate the effects and mechanisms of sennoside B on the growth,invasion of A549 cells and tumorigenesis of nude mice.Methods A549 non-small cell lung cancer cells were treated with sennoside B at doses of 0,5,10 and 20μM respectively.The cells were divided into four groups randomly according to the dose and then the follow-up experiment was proceeded.Brdu staining was used to detect the cell proliferation of each group,and Hoechst staining was used to detect apoptosis.Cell migration was detected by scratch test.Cell invasion was detected by Transwell assay.Protein expression levels of Ki67,PCNA,cl-caspase-3,cl-caspase-9,VEGF,N-cadherin and E-cadherin,as well as phosphorylation of STAT3 and ERK1/2 were detected by western blotting.Tumor-bearing mice model was established and the weight of tumor was tested.The expression of Ki67 and VEGF were detected by immunohistochemistry.Results Compared with the Control group,the result showed that the Brdu positive cells number,the invasion cells number in each sennoside B dosage group were significantly reduced(P<0.05),the apoptosis rate increased significantly(P<0.05),the rate of cells scratches healing(P<0.05),the protein levels of ki67,PCNA and VEGF,N-cadherin decreased obviously(P<0.05),the protein levels of clcaspase-3,cl-caspase-9,E-cadherin increased significantly(P<0.05),phosphorylation levels of STAT3 and ERK1/2 were significantly reduced(P<0.05),and the weight of tumor and the expression levels of Ki67 and VEGF were decreased in tumor-bearing mice model(P<0.05).Conclusions Sennoside B inhibits the phosphorylation of STAT3 and ERK1/2 and the growth of A549 cells.