华蟾素对非小细胞肺癌细胞株A549细胞增殖及PTEN/AKT/mTOR信号通路表达的影响

Effect of cinobufacin on the proliferation and expression of PTEN/AKT/mTOR signaling pathway in non-small cell lung cancer cell line A549

目的:观察华蟾素对肺癌细胞A549及PTEN/AKT/mTOR信号通路蛋白表达的影响。方法:在培养肺癌细胞株NCI-A549中分别加入不同浓度的华蟾素,应用细胞计数试剂盒-8检测24、48、72 h后细胞增殖活力,Ki67及EdU检测72 h细胞增殖活力,免疫印迹法检测磷脂酰肌醇-3激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白以及磷酸化第10号染色体缺失的磷酸酶和张力蛋白的同源基因(phosphorylated-phosphatase and tensin homolog deleted on chromosome ten,p-PTEN)蛋白表达。结果:0.2μg/mL华蟾素作用于A549细胞株48 h后显示明显抑制增殖作用(P<0.01),0.8μg/mL华蟾素处理12 h后均具有不同程度抑制增殖作用(P<0.05~P<0.01)。不同浓度华蟾素作用于细胞72 h后,p-PI3K、p-AKT、p-mTOR、p-S6、p-PTEN表达量均较对照组下降(P<0.05~P<0.01),而PI3K、AKT、mTOR、S6及PTEN表达量与对照组差异无统计学意义(P>0.05)。结论:华蟾素可能通过抑制PI3K/AKT/mTOR信号通路蛋白磷酸化,抑制肺癌细胞A549的增殖。

Objective:To observe the expression changes of PTEN/AKT/mTOR signaling pathway protein after the lung cancer cells A549 treated with cinobufacin.Methods:Different concentrations of cinobufacin were added to the cultured lung cancer cell line NCI-A549,and the cell proliferation activity was measured using cell counting kit-8 after 24,48 and 72 h.Ki67 and EdU were used to detect the proliferation of cells after 72 h of treatment with cinobufacin.Western blotting was used to detect the expression levels of protein kinase B/mammalian rapamycin target protein pathway protein(AKT/mTOR)and phosphorylated-phosphatase and tensin homolog deleted on chromosome ten(P-PTEN).Results:The proliferation of A549 cell line inhibited after treatment with 0.2μg/mL cinobufacin for 48 h(P<0.01),and the proliferation of A549 cell line was inhibited after treatment with 0.8μg/mL cinobufacin for 12 h(P<0.05 to P<0.01).Compared with the control group,the expression levels of p-PI3K,p-AKT,p-mTOR,p-S6 and p-PTEN in experimential group after 72 h of treatment with different concentrations of cinobufacin decreased(P<0.05 to P<0.01),while the differences of the expression levels of PI3K,AKT,mTOR,S6 and PTEN were not statistically significant between the control group and erperimantial group(P>0.05).Conclusions:Cinobufacin may inhibit the proliferation of lung cancer cells A549 by inhibiting the phosphorylation of PI3K/AKT/mTOR signaling pathway protein.