摘要
目的 探究黄芩素抑制喉癌细胞增殖及迁移的机制。方法 采用Hep-2细胞为研究对象。MTT法检测各组细胞增殖活力;划痕实验检测细胞迁移;流式细胞术检测细胞凋亡情况;Western blot检测各组细胞中Beclin-1、LC3II/I、p62及HDAC1的表达水平;RT-PCR检测细胞miR-449a的表达水平;生物信息学软件分析miR-449a与HDAC1的靶向结合位点,双荧光素酶报告基因实验鉴定;透射电子显微镜观察自噬小体数量。结果 黄芩素显著抑制了喉癌细胞的增殖和迁移,并诱导细胞凋亡;黄芩素增加了细胞内Beclin-1及LC3II的蛋白表达水平,减少了p62及LC3I的蛋白表达水平;黄芩素显著增加喉癌细胞miR-449a的表达水平,降低HDAC1的蛋白表达量;HDAC1与miR-449a存在结合作用;黄芩素联合miR-449a mimic处理喉癌细胞,可以更显著的抑制细胞增殖及HDAC1的蛋白表达水平,以及更为有效地上调miR-449a的相对表达量和诱导自噬。结论 黄芩素可以介导自噬经由miR-449a/HDAC1轴抑制喉癌细胞增殖及迁移。
Objective To explore the mechanism of baicalein inhibiting the proliferation and migration of laryngeal cancer cells.Methods Proliferation of Hep-2 laryngeal cancer cells was measured by MTT assays.Cell migration was assessed by a scratch assay.Apoptosis was detected by flow cytometry.Western blot was used to measure protein expression levels of Beclin-1,LC3II/I,p62 and HDAC1.The expression level of miR-449a was measured by RT-PCR.The binding sites of miR-449a and HDAC1 were analyzed by bioinformatics and identified by dual luciferase reporter assays.The number of autophagosomes was observed by projection microscopy.Results Baicalein significantly inhibited the proliferation and migration of laryngeal cancer cells,and significantly induced apoptosis.Baicalein increased the protein expression levels of intracellular autophagy-related genes Beclin-1 and LC3II,but decreased the protein expression levels of p62 and LC3I.Baicalein significantly increased the expression level of miR-449a in laryngeal cancer cells and decreased protein expression of HDAC1.Binding was observed between HDAC1 and miR-449a.Cotreatment of laryngeal cancer cells with Baicalein and an miR-449a mimic more significantly inhibited cell proliferation and HDAC1 protein expression,and more effectively upregulated miR-449a expression and induced autophagy.Conclusions Baicalein mediated autophagy and inhibited the proliferation and migration of laryngeal cancer cells via the miR-449a/HDAC1 axis.
作者
王剑
孙永东
周兴玮
刘雷
童兴科
陈龙
何娴
WANG Jian;SUN Yongdong;ZHOU Xingwei;LIU Lei;TONG Xingke;CHEN Long;HE Xian(Department of Otolaryngology,Head and Neck Surgery,Affiliated Hospital of Traditional Chinese Medicine,Southwest Medical University,Luzhou 646000,China;College of Integrated Chinese and Western Medicine,Southwest Medical University,Luzhou 646000)
出处
《中国比较医学杂志》
CAS
北大核心
2023年第10期45-53,共9页
Chinese Journal of Comparative Medicine
基金
西南医科大学附属中医医院联合项目课题(2020XYLH-024)。
