甘草甜素通过同源框基因D簇反义RNA1和微小RNA-30a-3p对鼻咽癌细胞的影响  

Effect of glycyrrhizin on nasopharyngeal carcinoma cells through HOXD-AS1/miR-30a-3p

作　　者：王晓晖 WANG Xiaohui(Department of Otolaryngology,Integrated Traditional Chinese and Western Medicine Hospital of Changping District,Beijing,102208,China)

机构地区：[1]北京市昌平区中西医结合医院耳鼻咽喉科,北京102208

出　　处：《中国耳鼻咽喉头颈外科》2022年第2期85-90,共6页Chinese Archives of Otolaryngology-Head and Neck Surgery

摘　　要：目的 探讨甘草甜素对鼻咽癌细胞的影响及分子机制。方法 鼻咽癌细胞C666-1随机分为对照组,不同浓度甘草甜素(25、50、100 μmol/L)组,si-NC组,si-HOXD-AS1组,甘草甜素+pcDNA-HOXD-AS1组,甘草甜素+anti-miR-30a-3p组;MTT法检测细胞增殖抑制率;平板克隆实验检测集落形成数;划痕实验检测细胞迁移距离;Transwell小室检测细胞侵袭数;蛋白质印迹(Western blot)法检测蛋白表达;实时荧光定量PCR(RT-qPCR)检测同源框基因D(homeobox D,HOXD)簇反义RNA 1(cluster antisense RNA1,HOXD-AS1)和微小RNA(microRNA,miRNA/miR)-30a-3p的表达水平;双荧光素酶报告实验检测HOXD-AS1和miR-30a-3p的靶向关系。结果 与对照组相比,不同浓度甘草甜素组C666-1细胞增殖抑制率升高,集落形成数减少,迁移距离缩短,侵袭细胞数减少,E-cadherin表达水平升高,N-cadherin表达水平降低,HOXD-AS1表达水平降低,miR-30a-3p表达水平升高,且呈浓度依赖性(P<0.05)。干扰HOXD-AS1后,细胞增殖抑制率升高,集落形成数减少,迁移距离缩短,侵袭细胞数减少,E-cadherin表达水平升高,N-cadherin表达水平降低(P<0.05)。HOXD-AS1靶向调控miR-30a-3p,过表达HOXD-AS1或抑制miR-30a-3p减弱了甘草甜素对C666-1细胞增殖、迁移侵袭的影响。结论 甘草甜素可能通过调控HOXD-AS1/miR-30a-3p轴抑制鼻咽癌细胞的增殖、迁移和侵袭。OBJECTIVE To explore the effect of glycyrrhizin on nasopharyngeal carcinoma cells and its molecular mechanism.METHODS Nasopharyngeal carcinoma cells C666-1 were randomly divided into control group,different concentrations of glycyrrhizin(25,50,100μmol/L) groups,si-NC group,si-HOXD-AS1 group,glycyrrhizin+pcDNA-HOXD-AS1group,glycyrrhizin+anti-miR-30a-3p group.Tetramethylazolium salt colorimetric method(MTT) was used to detect cell proliferation inhibition rate,plate cloning experiment was used to detect colony formation number,scratch test was used to detect cell migration distance,Transwell chamber was used to detect cell invasion number,Western blot was used to detect protein expression,and real-time fluorescent quantitative PCR(RT-qPCR) was used to detect the expression levels of HOXD-AS1 and miR-30a-3p.Dual luciferase reporter experiment was used to detect the targeting relationship between HOXD-AS1 and miR-30a-3p.RESULTS Compared with the control group,the C666-1 cell proliferation inhibition rate in the different concentrations of glycyrrhizin group was increased,the number of colonies formed was decreased,the migration distance was shortened,the number of invasive cells was decreased,the expression level of E-cadherin was increased,and the expression level of N-cadherin was decreased,the expression level of HOXD-AS1 was decreased,and the expression level of miR-30a-3p was increased,in a concentration-dependent manner(P<0.05).After interference with HOXD-AS1,the cell proliferation inhibition rate was increased,the number of colonies formed was decreased,the migration distance was shortened,the number of invasive cells was decreased,the expression level of E-cadherin was increased,and the expression level of N-cadherin was decreased(P<0.05).HOXD-AS1 targeted regulated the expression of miR-30a-3p.Overexpression of HOXD-AS1 or inhibition of miR-30a-3p weakened the effect of glycyrrhizin on the proliferation,migration and invasion of C666-1 cells.CONCLUSION Glycyrrhizin may inhibit the proliferation,migration a

关 键 词：鼻咽癌 细胞增殖 细胞迁移分析 甘草甜素 侵袭 

分 类 号：R285[医药卫生—中药学]