脱氢姜酮通过调控miR-223-3p抑制喉癌细胞Hep-2增殖、迁移及血管形成  

Dehydrogingerone inhibits proliferation,migration and angiogenesis of laryngeal cancer cells Hep-2 by regulating miR-223-3p

作　　者：吴俊 王文忠[1] 吴娟[1] 于宁静 孙哲[1] 周兰柱[1] WU Jun;WANG Wenzhong;WU Juan;YU Ningjing;SUN Zhe;ZHOU Lanzhu(Department of Otolaryngology Head and Neck Surgery,First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China)

机构地区：[1]蚌埠医学院第一附属医院耳鼻咽喉头颈外科,蚌埠233000

出　　处：《山西医科大学学报》2023年第4期446-453,共8页Journal of Shanxi Medical University

基　　金：安徽省高校自然科学研究重点项目(KJ2020A0590);蚌埠医学院自然科学重点项目(2021byzd061)。

摘　　要：目的探讨脱氢姜酮(DZG)对喉癌细胞Hep-2增殖、迁移及血管形成的影响及其相关机制。方法分别以不同浓度的DZG(0,75,150,300μmol/L)处理Hep-2细胞24 h,MTT法、Transwell、Western blot法、实时荧光定量PCR(RT-PCR)分别检测Hep-2细胞的存活率、迁移能力、迁移蛋白波形蛋白(Vimentin)、N-钙黏蛋白(N-cadherin)和血管形成蛋白血管内皮生长因子(VEGF)以及基质金属蛋白酶-2(MMP-2)相对表达水平和miR-223-3p mRNA相对表达量。将Hep-2细胞分别分为:①空白对照组(control组)、模拟物阴性对照组(miR-NC)、miR-223-3p组(转染miR-223-3p);②空白对照组(si-control组)、阴性对照组(si-NC)及miR-223-3p小干涉RNA组(si-miR-223-3p)。MTT法检测细胞增殖抑制能力,Transwell检测细胞迁移能力,Western blot检测细胞VGEF、MMP-2、Vimentin、N-cadherin蛋白相对表达量。结果0,75,150,300μmol/L的DZG培养Hep-2细胞,MTT结果显示,24 h细胞存活率随DZG浓度增加而下降(P<0.05);Transwell结果显示,细胞迁移能力随DZG浓度增加而下降(P<0.05);Western blot结果显示,VGEF、MMP-2、Vimentin、N-cadherin蛋白相对表达量随DZG浓度增加而降低(P<0.05);RT-PCR结果显示,miR-223-3p mRNA相对表达量随DZG浓度增加而增加(P<0.05)。将Hep-2细胞进行转染后,RT-PCR结果表明,miR-223-3p组Hep-2细胞miR-223-3p相对表达量较miR-NC组和control组明显升高,si-miR-223-3p组Hep-2细胞miR-223-3p相对表达量较si-NC组和si-control组明显降低(P<0.05)。MTT法结果显示,miR-223-3p组细胞存活率较miR-NC组明显降低(P<0.05)。不加DZG单独使用正常培养基培养验证miR-223-3p对Hep-2细胞存活率时发现,miR-223-3p组细胞存活率较miR-NC组和control组细胞明显下降,si-miR-223-3p组细胞存活率较si-NC组和si-control组明显升高(P<0.05);Transwell结果显示,miR-223-3p组细胞迁移率较miR-NC组和control组明显降低,si-miR-223-3p组细胞迁移率较si-NC组和si-control组明显升高(P<0.05)。Western blot结果�Objective To investigate the effects of dehydrogingerone(DZG)on proliferation,migration and angiogenesis of laryngeal cancer cells Hep-2 and its related mechanism.Methods Hep-2 cells were treated with DZG at different concentrations(0,75,150,300μmol/L)for 24 h.MTT method,Transwell,Western blot and RT-PCR were used to detect the survival rate of Hep-2 cells,the migration ability,the protein expression of Vimentin,N-cadherin,angioforming protein vascular endothelial growth factor(VEGF)and matrix metalloproteinase-2(MMP-2)and relative miR-223-3p mRNA level,respectively.Hep-2 cells were divided into:①blank control group(control group),negative analog control group(miR-NC),and miR-223-3p group(transfected with miR-223-3p);②blank control group(si-control group),negative control group(si-NC group)and miR-223-3p small interference RNA group(si-miR-223-3p group).MTT assay,Transwell assay and Western blot were used to detect the cell proliferation,the cell migration,and the relative expression levels of VGEF,MMP-2,Vimentin and N-cadherin.Results MTT and Transwell results showed that 24 h cell survival rate and the cell migration ability were statistically decreased by DZG in a concentration-dependent manner(P<0.05).Western blot results showed that the relative expression levels of VGEF,MMP-2,Vimentin and N-cadherin were decreased with the increase of DZG concentration(P<0.05).RT-PCR results showed that the mRNA relative expression level of miR-223-3p was increased in a concentration-dependent manner(P<0.05).RT-PCR results showed that the relative expression level of miR-223-3p in Hep-2 cells in miR-223-3p group was significantly higher than that in miR-NC group and control group and the relative expression level of miR-223-3p in Hep-2 cells in si-miR-223-3p group was significantly lower than that in si-NC group and si-control group(P<0.05).The results of MTT assay showed that the cell survival rate in miR-223-3p group was significantly lower than that in miR-NC group(P<0.05).Under normal culture medium without DZG,the

关 键 词：喉癌 miR-223-3p 增殖 侵袭 迁移 血管形成 

分 类 号：R739.65[医药卫生—肿瘤]