枞酸在顺铂耐药鼻咽癌细胞增殖、侵袭、迁移中的作用及分子机制
The role and the molecular mechanism of abietic acid in the proliferation,invasion and migration of cisplatin-resistant nasopharyngeal carcinoma cells
袁艳艳 1蓝苑钊 2崔毅 1侯明华 1唐金勇 1文浩杰2
作者信息
- 1. 郴州市第一人民医院耳鼻咽喉头颈外科(湖南郴州,423000)
- 2. 郴州市第一人民医院耳鼻咽喉头颈外科(湖南郴州,423000);南方医科大学第一临床学院
- 折叠
摘要
目的:探讨枞酸在顺铂耐药鼻咽癌细胞增殖、侵袭、迁移的作用及分子机制.方法:①药物浓度递增法构建顺铂耐药鼻咽癌C666/DDP细胞株.②枞酸对C666/DDP细胞增殖、侵袭及迁移的影响研究:CCK-8法、活性氧(ROS)及线粒体膜电位(MMP)水平检测及裸鼠皮下成瘤实验检测枞酸对C666/DDP细胞体外、体内增殖及凋亡能力的影响;Transwell法检测细胞侵袭和迁移能力.③Western blot及IHC法检测PI3K/AKT/mTOR通路相关蛋白的表达情况.结果:①顺铂对C666-1细胞毒性IC50约为25 μmol/L.RI=25μmol/L/4 μmol/L=6.25,获得耐药性,C666-1-DDP耐药株构建成功.②枞酸可促进C666/DDP细胞凋亡,抑制其增殖,且表现为G2/M期阻滞;Transwell显示枞酸抑制C666/DDP细胞迁移及侵袭能力,提高C666/DDP细胞ROS水平并使MMP降低.③动物实验显示枞酸在体内呈浓度梯度抑制顺铂耐药鼻咽癌的增殖,抑制PI3K/AKT/mTOR信号通路相关蛋白的表达.结论:枞酸可抑制顺铂耐药鼻咽癌细胞增殖、侵袭及迁移,其机制与抑制PI3K/AKT/mTOR信号通路有关.
Abstract
Objective:To investigate the effects and molecular mechanisms of abietic acid in the cell prolifera-tion,invasion and migration of cisplatin-resistant nasopharyngeal carcinoma cells.Methods:①Cisplatin-resistant C666/DDP cell line was constructed by increasing drug concentration method.②The effects of abietic acid on pro-liferation,invasion and migration of C666/DDP cells were investigated by CCK-8 method,reactive oxygen species(ROS)and mitochondrial membrane potential(MMP)level assay and subcutaneous tumorigenesis assay in nude mice to detect the effects of abietic acid on proliferation and apoptosis of C666/DDP cells in vitro and in vivo.The effect of abietic acid on the proliferation and apoptosis of C666/DDP cells in vitro and in vivo was measured by Tr-answell assay.③Western blot and IHC method to detect the expression of PI3K/AKT/mTOR pathway related proteins.Results:①The IC50 of cisplatin cytotoxicity to C666-1 was about 25 μmol/L.RI=25 µmol/L/4 μmol/L=6.25,resistance was obtained,and the C666-1-DDP resistant strain was successfully constructed.②Abietic acid promoted apoptosis and inhibited proliferation of C666/DDP cells,and showed G2/M phase block;transwell showed that abietic acid inhibited C666/DDP cell migration and invasion,increased ROS level of C666/DDP cells and decreased MMP.Transwell showed that abietic acid inhibited the migration and invasion ability of C666/DDP cells,increased the ROS level of C666/DDP cells and decreased MMP.③Animal experiments showed that abietic acid inhibited the proliferation of cisplatin-resistant nasopharyngeal carcinoma in vivo in a concentration gradient and suppressed the expression of PI3K/AKT/mTOR signaling pathway-related proteins.Conclusion:Abietic acid inhibits proliferation,invasion and migration of cisplatin-resistant nasopharyngeal carcinoma cells by a mechanism related to inhibition of PI3K/AKT/mTOR signaling pathway.
关键词
枞酸/鼻咽肿瘤/顺铂耐药/分子机制Key words
abietic acid/nasopharyngeal carcinoma/cisplatin resistance/molecular mechanism引用本文复制引用
基金项目
湖南省自然科学基金区域联合基金(2023JJ50380)
郴州市科技计划(zdyf201936)
郴州市第一人民医院优青项目(N2020-2)
出版年
2024
NETL
NSTL
万方数据