Kaempferide is an O-methylated flavonol that has received much attention due to its various biological activities. In this study, we explored the underlying mechanisms of kaempferide in human lung cancer A549 cells. The Cell Counting Kit-8 (CCK-8) assay, Hoechst 33342/propidium iodide double staining, flow cytometry, scratch wound healing assay, and Western blot analysis were used to measure cell apoptosis, the cell cycle, reactive oxygen species (ROS) levels, and cell migration of human lung cancer cells. Kaempferide significantly inhibited human lung cancer cell proliferation, and its toxic effects on normal cells were significantly lower than those of 5-fluorouracil. Kaempferide induced A549 cell apoptosis by decreasing the mitochondrial membrane potential and the expression level of B-cell lymphoma 2, and by increasing the expression levels of Bcl-2-associated X protein and caspase-3. It also regulated mitogen-activated protein kinase (MAPK), signal transducer and activator of transcription 3 (STAT3), and nuclear factor kappa B (NF-κB) signaling pathways by increasing the expression levels of phosphorylated c-Jun N-terminal kinase, p-p38, I kappa B, and by decreasing the expression levels of phosphorylated extracellular signal-regulated kinase, p-STAT3, and NF-κB. Kaempferide induced cell cycle arrest in the G0/G1 phase in A549 cells by downregulating the expression levels of p-AKT, cyclin D1, and cyclin-dependent kinase 2. Furthermore, kaempferide blocked A549 cell migration by downregulating the expression levels of transforming growth factor beta 1 (TGF-β1), p-β-catenin, p-glycogen synthase kinase 3 beta, N-cadherin, and vimentin, and by upregulating the expression level of E-cadherin. Kaempferide enhanced the accumulation of ROS, and N-acetyl-l-cysteine (a ROS inhibitor) decreased the regulation of MAPK, NF-κB, AKT, and TGF-β signaling pathways by kaempferide, inhibited cell apoptosis, and reversed cell cycle arrest. Our results showed that kaempferide induced apoptosis via ROS-mediated MAPK, NF-κB, AKT, and TGF-β signaling pathways in A549 cells. Thus, kaempferide may be a novel drug candidate for lung cancer.

Kaempferide是一个O-甲基化黄酮醇由于其多种生物活性而备受关注。在这项研究中，我们探索了卡恩培利特在人类肺癌A549细胞中的潜在机制。细胞计数试剂盒8（CCK-8）测定，Hoechst 33342 /碘化丙啶双染色，流式细胞仪，刮擦伤口愈合测定和蛋白质印迹分析用于测量细胞凋亡，细胞周期，活性氧（ROS）肺癌细胞的水平和细胞迁移。Kaempferide显着抑制人肺癌细胞的增殖，其对正常细胞的毒性作用明显低于5-氟尿嘧啶。Kaempferide通过降低线粒体膜电位和B细胞淋巴瘤2的表达水平来诱导A549细胞凋亡2 并且通过增加Bcl-2相关X蛋白和caspase-3的表达水平。它还通过增加磷酸化的c-Jun N末端激酶的表达水平来调节丝裂原激活的蛋白激酶（MAPK），信号转导和转录激活因子3（STAT3）以及核因子κB（NF-κB）信号通路， p-p38，IκB，并通过降低磷酸化的细胞外信号调节激酶，p-STAT3和NF-κB的表达水平。Kaempferide通过下调p-AKT，cyclin D1和cyclin依赖性激酶2的表达水平，诱导A549细胞G0 / G1期细胞周期停滞。此外，kaempferide通过下调转化生长因子的表达水平来阻断A549细胞迁移。 beta 1（TGF-β1），p-β-catenin，p-糖原合酶激酶3 beta，N-cadherin和波形蛋白，并通过上调E-cadherin的表达水平。Kaempferide增强了ROS的积累，并且N-乙酰基-1-半胱氨酸（一种ROS抑制剂）降低了Kaempferide对MAPK，NF-κB，AKT和TGF-β信号通路的调节，抑制了细胞凋亡，并逆转了细胞周期停滞。我们的结果表明，kaempferide通过ROS介导的MAPK，NF-κB，AKT和TGF-β信号通路诱导A549细胞凋亡。因此，kaempferide可能是肺癌的一种新的候选药物。