肺金生方调控TGF-β/Smad信号通路抗肺癌侵袭

Effects of Feijinsheng Formula on the invasion of lung cancer by regulating the TGF-β/Smad signal pathway

目的:探讨肺金生方在体外抗肺癌转移的作用。方法:以肺金生方冻干粉作用于肺癌A549细胞,普通倒置显微镜观察结晶紫染色细胞的形态,CCK8法检测细胞增殖情况,划痕试验观察细胞迁移,ELISA、Western Blot法检测转化生长因子β(TGF-β)/Smad信号通路关键分子的表达。结果:梯度浓度的肺金生方冻干粉作用24h后,1、2mg/mL的药物可显著抑制A549细胞的活力,其细胞存活率较对照组显著降低(P<0.01);细胞形态由规则的偏圆形转变为不规则形,细胞数量减少,聚集情况减少;A549细胞迁移率分别为0mg/mL组(72.8±13.3)%、0.5mg/mL组(51.7±9.9)%、1mg/mL组(37.9±9.4)%、2mg/mL组(18.5±13.4)%;1mg/mL肺金生方下调了TGF-β1、Smad2/3和上调了Smad7表达,与空白组比较差异有统计学意义(P<0.05,P<0.01);2mg/mL肺金生方下调了TGF-β1、p-Smad2/3的表达,上调了Smad7的表达,与0mg/mL组比较,有显著差异(P<0.05)。结论:肺金生方能通过调控TGF-β/Smad信号通路达到抗肺癌转移的作用。

Objective:To study the effects of Feijinsheng Formula on anti-tumor metastasis of lung cancer cell lines in vitro.Methods:The lyophilized powder of Feijinsheng Formula was applied to lung cancer A549 cells.The morphology of crystal violet staining cells was observed by an inverted microscope.The cell proliferation was detected by CCK8 method.Cell migration was observed by the scratch test.The expression of key molecules of the TGF-β/Smad signaling pathway was detected by ELISA and Western Blot.Results:After being treated by the lyophilized powder of Feijinsheng Formula with gradient concentration for 24 hours,1 mg/mL and 2 mg/mL drugs could significantly inhibit the viability of A549 cells,and the cell survival rate was significantly lower than that of the control group(P<0.01).The circular shape was changed into an irregular shape,the number of cells decreased,and the aggregation decreased.The migration rate of A549 cells was(72.8±13.3)%in the control group,(51.7±9.9)%in the 0.5 mg/mL group,(37.9±9.4)%in the 1 mg/mL group and(18.5±13.4)%in the 2 mg/mL group(P<0.05,P<0.01);1 mg/mL Feijinsheng Formula down-regulated TGF-β1,Smad2/3 and up-regulated Smad7 expression.There was a difference compared with the blank group(P<0.05).The expression of TGF-β1 and p-Smad2/3 was down-regulated and the protein expression of Smad7 was up-regulated.There was significant difference compared with 0 mg/mL group(P<0.05).Conclusion:Feijinsheng Formula can regulate the TGF-β/Smad signal pathway to anti lung cancer metastasis.