背景:黄芪甲苷(ASIII),一种来源于中药黄芪的类皂苷代谢产物,已被证明可以有效治疗癌症和心力衰竭,和各种消化系统疾病。然而,其治疗非小细胞肺癌(NSCLC)的分子机制尚不清楚。
方法:用不同浓度的ASⅢ处理人肺癌A549细胞、NCI-H460细胞和正常人肺上皮细胞BEAS-2B。CCK-8和EdU染色用于确定ASIII的体外抗增殖作用。在用指定浓度的ASIII处理的A549细胞上进行定量蛋白质组学分析,免疫印迹法检测细胞凋亡相关蛋白的表达水平。
结果:ASIII治疗可显著抑制A549和H460细胞的增殖和增加细胞凋亡,并调节与凋亡和代谢相关的功能信号通路。在分子水平上,ASIII促进了ANXA1表达的降低(p<0.01),与增加水平的切割半胱天冬酶3和PARP1。此外,ASIII治疗显著降低了LC3-I/LC3-II比率。体外实验结果表明,ASⅢ通过下调P38、JNK、和AKT(p<0.01),抑制Bcl-2的表达(p<0.01),上调Bax的表达(p<0.01)。
结论:这些发现提供了ASIII治疗诱导NSCLC细胞凋亡的机制,这可以部分地通过调节P38、ERK和mTOR信号通路来实现。
BACKGROUND: Astragaloside III (AS III), a saponin-like metabolite derived from the traditional Chinese medicine Astragali Radix, has been shown to be effective in the treatment of cancer and heart failure, and a variety of digestive disorders. However, its molecular mechanism in the treatment of non-small cell lung cancer (NSCLC) is unknown.
METHODS: Human lung cancer A549 cells and NCI-H460 cells and a normal human lung epithelial cell BEAS-2B were treated with different concentrations of AS III. CCK-8 and EdU staining were used to determine the anti-proliferative effects of AS III in vitro. Quantitative proteomic analysis was performed on A549 cells treated with the indicated concentrations of AS III, and the expression levels of apoptosis-related proteins were examined by Western blotting.
RESULTS: AS III treatment significantly inhibited proliferation and increased apoptosis in A549 and H460 cells and modulated functional signaling pathways associated with apoptosis and metabolism. At the molecular level, AS III promoted a reduction in the expression of ANXA1 (p < 0.01), with increased levels of cleaved Caspase 3 and PARP 1. In addition, AS III treatment significantly decreased the LC3-I/LC3-II ratio. The results of experiment in vitro showed that AS III promoted NSCLC apoptosis by down-regulating the phosphorylation levels of P38, JNK, and AKT (p < 0.01), inhibiting the expression of Bcl-2 (p < 0.01), and up-regulating the expression of Bax (p < 0.01).
CONCLUSIONS: These findings provide a mechanism whereby AS III treatment induces apoptosis in NSCLC cells, which may be achieved in part via modulation of the P38, ERK and mTOR signaling pathways.