摘要
目的:探究甘草甜素(Gly)对喉癌Hep-2细胞侵袭和迁移的影响及作用机制。方法:体外培养正常人喉黏膜上皮细胞和喉癌Hep-2细胞,实时荧光定量PCR(RT-qPCR)法和蛋白印迹实验检测微小RNA-205-5p(miR-205-5p)、沉默信息调节因子2相关酶3(SIRT3)mRNA和蛋白表达水平。将喉癌Hep-2细胞分为对照组、Gly低浓度组、Gly中浓度组、Gly高浓度组、Gly高浓度+空载质粒组、Gly高浓度+miR-205-5p抑制剂组。各组给予相应浓度的Gly干预或转染对应质粒培养48 h。Transwell实验和划痕实验分别检测各组Hep-2细胞侵袭和迁移能力。RT-qPCR法和蛋白印迹实验检测各组Hep-2细胞miR-205-5p、SIRT3 mRNA和蛋白表达水平。双荧光素酶报告基因实验分析miR-205-5p与SIRT3的靶向关系。结果:与人喉黏膜上皮细胞比较,喉癌Hep-2细胞miR-205-5p水平降低,SIRT3 mRNA和蛋白水平升高(均P<0.05)。与对照组比较,Gly低、中、高浓度组穿膜细胞数和划痕愈合率、SIRT3 mRNA和蛋白水平依次降低,miR-205-5p水平依次升高(均P<0.05)。与Gly高浓度组和Gly高浓度+空载质粒组比较,Gly高浓度+miR-205-5p抑制剂组穿膜细胞数和划痕愈合率、SIRT3 mRNA和蛋白水平升高,miR-205-5p水平降低(均P<0.05)。miR-205-5p可靶向调控SIRT3表达。结论:Gly能够抑制Hep-2细胞侵袭和迁移,其机制可能与上调miR-205-5p表达,进而靶向下调SIRT3表达有关。
Objective:To explore the effect of glycyrrhizin(Gly)on invasion and migration of laryngeal carcinoma Hep-2 cells and its mechanism.Methods: Normal human laryngeal mucosal epithelial cells and laryngeal cancer Hep-2 cells were cultured in vitro,and the microRNA-205-5p(miR-205-5p),silent mating type information regulation 2 homolog-3(SIRT3)mRNA and protein levels were detected by real-time fluorescent quantitative PCR(RT-qPCR)and western blot assay.Laryngeal cancer Hep-2 cells were divided into control group,Gly low concentration group,Gly medium concentration group,Gly high concentration group,Gly high concentration+empty plasmid group,Gly high concentration+miR-205-5p inhibitor group.Each group was given Gly intervention with corresponding concentration or transfected with corresponding plasmid for 48 h.The invasion and migration ability of Hep-2 cells in each group were detected by Transwell assay and scratch assay respectively.The miR-205-5p,SIRT3 mRNA and protein levels of Hep-2 cells in each group were detected by RT-qPCR and Western blot.The targeting relationship between miR-205-5p and SIRT3 was analyzed by double luciferase reporter gene assay.Results: Compared with human laryngeal mucosal epithelial cells,the levels of miR-205-5p in laryngeal cancer Hep-2 cells were decreased,and the levels of SIRT3 mRNA and protein were increased(all P<0.05).Compared with control group,the number of transmembrane cells,scratch healing rate,SIRT3 mRNA and protein levels in Gly low,medium and high concentration groups were decreased successively,and the levels of miR-205-5p were increased successively(all P<0.05).Compared with Gly high concentration group and Gly high concentration+empty plasmid group,the number of transmembrane cells,scratch healing rate,SIRT3 mRNA and protein levels in Gly high concentration+miR-205-5p inhibitor group were increased,and the level of miR-205-5p was decreased(all P<0.05).miR-205-5p could target the regulation of SIRT3 expression.Conclusion:Gly can inhibit the invasion and migration of Hep-2 cells,and the mechanism may be related to the up-regulation of miR-205-5p expression and the targeted down-regulation of SIRT3 expression.
作者
要兆旭
马海滨
刘琳
赵倩
巩慧
孙凯丽
秦隆朝
YAO Zhaoxu;MA Haibin;LIU Lin;ZHAO Qian;GONG Hui;SUN Kaili;QIN Longchao(Department of Otolaryngology,Handan Central Hospital,Handan 056002,China)
出处
《陕西医学杂志》
CAS
2024年第5期604-609,共6页
Shaanxi Medical Journal
基金
河北省医学科学研究课题(20230468)。
