新型荜茇酰胺衍生物C12阻滞A549细胞周期、诱导细胞凋亡和自噬

Novel Piperlongumine Derivative C12 Induces Cell Cycle Arrest,Apoptosis,and Autophagy in A549 Cells

该研究旨在探讨荜茇酰胺衍生物C12对非小细胞肺癌(NSCLC)A549的体外抑制活性及其作用机制。采用四甲基偶氮唑蓝(MTT)法和克隆形成实验检测C12对细胞增殖的影响;划痕实验和Transwell实验检测C12对细胞迁移和侵袭的影响;流式细胞术检测对细胞周期和凋亡的影响;采用转录组测序检测C12处理后的差异表达基因;Western blot验证C12对相关蛋白表达变化的影响。结果显示,C12能有效抑制A549的增殖、迁移和侵袭,阻滞细胞周期于G2/M期,并通过调控Bax和Bcl-2的表达水平诱导细胞凋亡;其调控基因主要存在于细胞染色体组分中,显著富集于催化活性和DNA结合相关的分子功能、细胞周期生物学过程和MAPK、PI3K/Akt信号通路;C12能上调细胞内的ROS水平,通过上调p-JNK、p-Erk1/2和p-p38的表达来激活MAPK信号通路诱导细胞凋亡;同时,C12通过下调p-Akt和p-mTOR,上调LC3-II诱导细胞自噬。总之,C12在A549中可以激活MAPK信号通路诱导细胞凋亡和抑制PI3K/Akt/m-TOR信号通路诱导细胞自噬。

This study aimed to investigate the inhibitory effect of Piperlongumine derivative C12 on NSCLC(non-small cell lung cancer)cell line A549 and its mechanism in vitro.MTT assay and colony formation assay were used to detect the effect of C12 on cell proliferation.Scratch assay and Transwell assay were used to detect the effects of C12 on cell migration and invasion.Cell cycle and apoptosis were detected by flow cytometry.Transcriptome sequencing was used to predict DEGs(differentially expressed genes)after C12 treatment.Western blot was used to verify the effect of C12 on the expression of related proteins.The results showed that C12 could effectively inhibit the proliferation,migration,and invasion of A549 cells,arrest cell cycle at G2/M phase and induce cell apoptosis by regulating the expression of Bax and Bcl-2.The regulatory genes were mainly present in the chromosomal component and were significantly enriched in molecular functions related to catalytic activity and DNA binding,cell cycle biological processes,and MAPK and PI3K/Akt signaling pathways.C12 could increase intracellular ROS level and activate MAPK signaling pathway to induce cell apoptosis by up-regulating the expression of p-JNK,p-Erk1/2 and p-p38.At the same time,C12 induced autophagy by down-regulating p-Akt and p-mTOR and up-regulating LC3-II.In conclusion,C12 could activate MAPK signaling pathway to induce apoptosis and inhibit PI3K/Akt/m-TOR signaling pathway to induce autophagy in A549 cells.