荜茇酰胺衍生物C12对人非小细胞肺癌H1299细胞的抗肿瘤作用及其机制研究  

The antitumor activity and mechanisms of piperlongumine derivative C12 on human non-small cell lung cancer H1299 cells

作　　者：龙海洮 雷雪 陈佳宜 孟娇[3] 邵利辉 李洙锐 陈丹萍[1] 王贞超 周玥[1,2] 李成朋 LONG Hai-tao;LEI Xue;CHEN Jia-yi;MENG Jiao;SHAO Li-hui;LI Zhu-rui;CHEN Dan-ping;WANG Zhen-chao;ZHOU Yue;LI Cheng-peng(College of Pharmacy,Guizhou University,Guiyang 550025,China;Guizhou Engineering Laboratory for Synthetic Drugs,Guizhou University,Guiyang 550025,China;National Key Laboratory of Green Pesticide,Key Laboratory of Green Pesticide and Agricultural Bioengineering,Ministry of Education,Center for R&D of Fine Chemicals of Guizhou University,Guiyang 550025,China)

机构地区：[1]贵州大学药学院,贵州贵阳550025 [2]贵州大学,贵州省合成药物工程实验室,贵州贵阳550025 [3]绿色农药国家重点实验室,绿色农药与农业生物工程教育部重点实验室,贵州大学精细化学品研发中心,贵州贵阳550025

出　　处：《药学学报》2024年第10期2773-2781,共9页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(22007022,32360689,22364008,32260694,21867004);贵州省自然科学基金(ZZK[2021]034);贵州省教育厅拔尖科技人才计划(2022075)。

摘　　要：(E)-1-(4-(3-(5-氯-6-氧代-3,6-二氢吡啶-1(2H)-基)-3-氧代丙基-1-烯-1-基)苯基)-3-(4-氟苯基)脲(以下简称C12)是课题组前期合成的一个新型荜茇酰胺衍生物。本研究以人非小细胞肺癌H1299细胞为研究对象,探讨C12对人非小细胞肺癌的体外抗肿瘤作用及其作用机制。采用噻唑蓝(methyl thiazolyl tetrazolium、MTT)法、划痕实验、平板克隆实验和Transwell细胞侵袭实验检测C12对H1299细胞增殖、迁移和侵袭的影响;通过流式细胞术检测C12对H1299细胞周期、活性氧(reactive oxygen species,ROS)的产生、线粒体膜电位(mitochondrial membrane potential,MMP)和细胞凋亡的影响;通过蛋白印迹实验检测p21、Cyclin B1、CDK1、Bax、Bcl-2、JNK、p-JNK、Erk1/2、p-Erk1/2、p38和p-p38的表达,探讨C12的抗肿瘤作用机制。结果显示,C12呈时间和浓度依赖性地抑制H1299的增殖、迁移和侵袭;流式细胞结果显示,C12能阻滞H1299细胞周期于G2/M期,升高ROS水平,降低MMP并诱导细胞凋亡;蛋白印迹结果显示,C12通过下调Cyclin B1和CDK1蛋白表达将H1299细胞阻滞于G2/M期,上调Bax/Bcl-2水平诱导细胞凋亡,上调MAPK通路中p-JNK、p-Erk1/2和p-p38的表达。综上所述,C12能够显著抑制H1299的增殖、迁移和侵袭,并诱导细胞周期阻滞和细胞凋亡,其机制可能与激活MAPK信号通路有关。The compound(E)-1-(4-(3-(5-chloro-6-oxo-3,6-dihydropyridin-1(2H)-yl)-3-oxo-propyl-1-ene-1-yl)phenyl)-3-(4-fluorophenyl)urea(C12),a novel derivative of piperlongumine previously synthesized by our research group,was investigated in this study to examine its effects on human non-small cell lung cancer cell line H1299 in vitro and elucidate its potential mechanism of action.The impact of C12 on the proliferation,migration,and invasion abilities of H1299 cells were assessed using methyl thiazolyl tetrazolium(MTT)assay,wound healing assay,cloning formation assay,and Transwell assay.Flow cytometry was employed to evaluate the influence of C12 on cell cycle progression,reactive oxygen species(ROS)production,mitochondrial membrane potential(MMP),and apoptosis induction in H1299 cells.Western blot analysis was conducted to investigate the expression levels of p21,Cyclin B1,CDK1,Bax,Bcl-2,JNK,p-JNK,Erk1/2,p-Erk1/2,p38 and p-p38 proteins for exploring the anti-tumor mechanism underlying C12's actions.The results demonstrated that C12 exerted inhibitory effects on the proliferation,migration,and invasion capacities of H1299 cells in a time-dependent and concentration-dependent manner.Moreover,C12 induced G2/M phase arrest in the cell cycle,reduced MMP levels,elevated ROS production,and triggered apoptotic processes.Flow cytometry analysis revealed that C12 downregulated Cyclin B1 and CDK1 protein expressions,resulting in G2/M phase arrest.C12 also upregulated Bax/Bcl-2 ratio,promoting apoptosis.Furthermore,C12 activated MAPK signaling pathway by enhancing phosphorylation levels of JNK,Erk1/2,and p38 proteins.In conclusion,C12 significantly suppressed proliferation,migration,and invasion capabilities while inducing cell cycle arrest and apoptosis in H1299 cells.These effects may be attributed to activation of the MAPK signaling pathway.

关 键 词：荜茇酰胺衍生物 H1299细胞 增殖 凋亡 MAPK信号通路 

分 类 号：R966[医药卫生—药理学]