苦参碱通过PI3K/Akt信号通路诱导肺癌A549细胞凋亡机制研究

Study on the apoptosis mechanism of lung cancer A549 cells induced by matrine through PI3K/Akt signaling pathway

目的探究苦参碱通过PI3K/Akt信号通路诱导肺癌A549细胞凋亡机制。方法对人肺癌A549细胞进行培养,培养后分为对照组、低浓度组、中浓度组以及高浓度组,对照组中加入生理盐水,分别向低浓度组、中浓度组以及高浓度组,加入浓度为30 mg/L、60 mg/L、120 mg/L的苦参碱,作用48小时后采用流式细胞仪及MTT检测法检测A549细胞凋亡情况,计算生长抑制率,并采用Western Blotting和real time PCR检测PI3K、p-AKT蛋白以及PI3K、AKT mRNA表达水平,比较四组之间的差异。结果高浓度组、中浓度组、低浓度组生长抑制率和凋亡率显著高于对照组(P<0.05),且随着浓度的升高生长抑制率和凋亡率显著升高(P<0.05);高浓度组、中浓度组、低浓度组PI3K、AKT mRNA及PI3K、p-AKT蛋白表达显著低于对照组(P<0.05),且随着浓度的升高PI3K、AKT mRNA及PI3K、p-AKT蛋白表达水平显著降低(P<0.05)。结论苦参碱可诱导肺癌A549细胞凋亡,推测其作用机制与PI3K/Akt信号通路有关。

Objective To explore the apoptosis mechanism of lung cancer A549 cells induced by matrine through PI3K/Akt signaling pathway. Methods Human lung cancer A549 cells were cultured,which were divided into control group,low concentration group,middle concentration group and high concentration group. The cells in control group were given normal saline,and the low concentration group,middle concentration group and high concentration group were respectively given matrine with concentrations of 30 mg/L,60 mg/L,120 mg/L,and the apoptosis situations of lung cancer A549 cells were detected by flow cytometry and MTT detection,and the growth inhibition rates were calculated,then the PI3K,p-AKT protein and PI3K,AKT mRNA levels were detected by Western Blotting and real time PCR,the differences between the four groups were compared. Results The growth inhibition rates and apoptosis rates of high concentration group,middle concentration group and low concentration group were significantly higher than those in the control group（P〈0.05）,which were significant increased with the increasing of concentrations（P〈0.05）. The levels of PI3K,AKT mRNA and PI3K,p-AKT protein in the high concentration group,middle concentration group and low concentration group were significantly lower than those in the control group（P〈0.05）,which were significantly decreased with the increasing of concentrations（P〈0.05）. Conclusion The matrine can induce the apoptosis of lung cancer A549 cells,and the action mechanism may relate to the PI3K/Akt signaling pathway.