Abstract：
In the current work, grifolin was obtained from the twigs and leaves of Daphne genkwa for the first time and displayed significant growth inhibition against human lung carcinoma A549 cells. Subsequent in vitro antitumor evaluation revealed that grifolin could induce remarkable cell apoptosis and G0/G1 phase arrest, as well as block cell migration and invasion. In addition, grifolin also disrupted cellular energy metabolism by inducing reactive oxygen species, reducing adenosine triphosphate and mitochondrial membrane potential, and damaging DNA synthesis. Further RNA-seq analysis demonstrated that treatment of grifolin on A549 cells led to gene enrichment in MAPK, PI3K/Akt and NF-κB signaling pathways, all of which were inhibited by grifolin according to immunoblotting experiments. Further mechanistical studies disclosed that the expression of a key upstream protein KRAS was also blocked, and the cell death triggered by grifolin could be rescued by a RAS activator ML-099. Moreover, pretreatment of ML-099 on A549 cells could reverse the grifolin-induced downregulation of key proteins in the three aforementioned pathways. These findings indicate that grifolin could induce cell death in A549 cell line by inhibiting KRAS-mediated multiple signaling pathways.
摘要：
在目前的工作中,grifolin首次从Daphnegenkwa的树枝和叶片中获得，并显示出对人类肺癌A549细胞的显着生长抑制。随后的体外抗肿瘤评估表明，grifolin可以诱导显著的细胞凋亡和G0/G1期阻滞，以及阻断细胞迁移和入侵。此外,grifolin还通过诱导活性氧破坏细胞能量代谢，减少三磷酸腺苷和线粒体膜电位，破坏DNA合成.进一步的RNA-seq分析表明，grifolin对A549细胞的处理导致MAPK基因富集，PI3K/Akt和NF-kB信号通路,根据免疫印迹实验，所有这些都被grifolin抑制。进一步的机制研究表明，关键上游蛋白KRAS的表达也被阻断，并且由grifolin触发的细胞死亡可以通过RAS激活剂ML-099来挽救。此外，ML-099对A549细胞的预处理可以逆转grifolin诱导的上述三种途径中关键蛋白的下调。这些发现表明grifolin可以通过抑制KRAS介导的多种信号通路诱导A549细胞系细胞死亡。
