摘要
目的:探讨羽扇豆醇对人鼻咽癌CNE2细胞增殖、迁移、侵袭、凋亡、周期及对放、化疗敏感性的影响,同时探讨其凋亡机制。方法:CCK-8法检测羽扇豆醇对CNE2细胞增殖的影响以及联合顺铂对CNE2细胞的增殖抑制作用有无增强;细胞划痕、Transwell实验、流式细胞术分别用于检测羽扇豆醇对CNE2细胞迁移、侵袭、凋亡及周期的影响;平板细胞克隆形成实验检测羽扇豆醇联合X线放射处理对CNE2细胞克隆形成能力的影响;Western blot法检测羽扇豆醇对CNE2细胞凋亡信号通路中关键蛋白Bcl-2、Bax及Cleaved caspase-3的影响,探究羽扇豆醇的抗肿瘤机制。结果:羽扇豆醇呈药物浓度和时间依赖性地抑制CNE2细胞的增殖,选取羽扇豆醇40、60、80μmol/L用于后续实验。羽扇豆醇显著降低了CNE2细胞的迁移、侵袭能力,出现典型的凋亡特征,G1期延长,S期、G2期缩短,提高了顺铂或X线放射对CNE2细胞增殖及克隆形成的抑制能力,同时引起CNE2细胞中Bax、Cleaved caspase-3蛋白表达升高,Bcl-2蛋白表达下调(所有结果均P<0.001或P<0.0001)。结论:羽扇豆醇抑制CNE2细胞的增殖、迁移、侵袭,诱导CNE2细胞的凋亡,引起细胞周期阻滞在G1期,同时可提高顺铂及X射线对CNE2细胞的敏感性。证实了羽扇豆醇诱导CNE2细胞凋亡可能与调控凋亡信号通路中凋亡蛋白Bax、凋亡启动因子Cleaved caspase-3升高及抗凋亡蛋白Bcl-2降低有关。
Objective:To investigate the effects of lupeol on the proliferation,migration,invasion,apoptosis,cycle,sensitivity to radiotherapy and chemotherapy of human nasopharyngeal carcinoma CNE2 cells,and to explore the mechanism of apoptosis.Methods:CCK-8 method was used to determine the effect of lupeol on the proliferation of CNE2 cells and whether the inhibitory effect of lupeol combined with cisplatin on CNE2 cells was enhanced.Cell scratch test,Transwell test and Flow cytometry were used to determine the effects of lupeol on migration,invasion,ap⁃optosis and cell cycle of CNE2 cells.The effect of lupeol combined with X-ray irradiation on the colony forming abil⁃ity of CNE2 cells was measured by plate cell clone formation test.The effects of lupeol on Bcl-2,Bax and Cleaved caspase-3 in CNE2 cell apoptosis signaling pathway were determined by Western blot to explore the anti-tumor mechanism of lupeol.Results:Lupeol inhibited the proliferation of CNE2 cells in a concentration and time-depend⁃ent manner.Lupeol 40,60,80μmol/L were selected for subsequent experiments.Lupeol significantly reduced the mi⁃gration and invasion ability of CNE2 cells,and presented typical apoptotic characteristics.The cell cycle of G1 phase was prolonged,S phase and G2 phase were shortened.Lupeol enhanced the inhibition of cisplatin or X-ray radiation on proliferation and clone formation of CNE2 cells.At the same time,the expression of Bax and Cleaved caspase-3 proteins in CNE2 cells were increased,and the expression of Bcl-2 protein was down regulated(All results were P<0.001 or P<0.0001).Conclusion:Lupeol inhibited the proliferation,migration and invasion of CNE2 cells,induced apoptosis of CNE2 cells,and caused cell cycle arrest in G1 phase.At the same time,it could improve the sensitivity of cisplatin and X-ray to CNE2 cells.It was confirmed that the apoptosis of CNE2 cells induced by lupeol may be relat⁃ed to the increase of apoptosis proteins Bax and Cleaved caspase-3 and the decrease ofanti-apoptosis protein Bcl-2.
作者
王安生
李源泉
冯燚源
杨晶
吴春成
邓世山
杨洪斌
WANG Ansheng;LI Yuanquan;FENG Yiyuan;YANG Jing;WU Chuncheng;DENG Shishan;YANG Hongbin(Department of Otorhinolaryngology Head and Neck Surgery,Affiliated Hospital of North Sichuan Medical College,Sichuan Nanchong 637000,China;North Sichuan Medical College,Sichuan Nanchong 637000,China;Institute of Medical Imaging,North Sichuan Medi-cal College,Sichuan Nanchong 637000,China)
出处
《现代肿瘤医学》
CAS
北大核心
2023年第4期618-625,共8页
Journal of Modern Oncology
基金
川北医学院科研发展计划项目(编号:CBY21-QA28)。
关键词
羽扇豆醇
鼻咽癌
放射
顺铂
凋亡
lupeol
nasopharyngeal carcinoma
radiation
cisplatin
apoptosis
