Southwest Medical University Hospital of Traditional Chinese Medicine
Joint project topic at the Hospital of Traditional Chinese Medicine of Southwest Medical University
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；WB检测各组细胞中Beclin-1、LC3I/II、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Beclin1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miR449a/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: Hep-2 cells were used as the research object. The cell proliferation activity of each group was detected by MTT method, cell migration was detected by scratch assay, cell apoptosis was detected by flow cytometry, western blot was used to detect the expression levels of Beclin-1, LC3II/I, p62 and HDAC1 in laryngeal cancer cells in each group, RT-PCR detection of the expression levels of miR-449a in each group of cells. The targeted binding sites of miR-449a and HDAC1 were analyzed by bioinformatics software and identified by dual luciferase reporter gene experiments. 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 Beclin1 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 the protein expression of HDAC1. There is a binding effect between HDAC1 and miR-449a. Baicalein combined with miR-449a mimic to treat laryngeal cancer cells can more significantly inhibit cell proliferation and HDAC1 protein expression, as well as more effectively up-regulate the relative expression of miR-449a and induce autophagy. Conclusion: Baicalein can mediate autophagy and inhibit the proliferation and migration of laryngeal cancer cells via the miR449a/HDAC1 axis.