miR-21对人口腔癌细胞株HSQ-89增殖、侵袭与迁移的影响实验研究  

Effect of miR-21 on proliferation,invasion and migration of human oral cancer cell line HSQ-89

作　　者：韦敏 顾春梅[1] 管燕华[1] 王育新[1] WEI Min;GU Chunmei;GUAN Yanhua;WANG Yuxin(Department of Oral and Maxillofacial Surgery,Stomatological Hospital Affiliated to Medical School of Nanjing University,Nanjing 210008,China)

机构地区：[1]南京大学医学院附属口腔医院口腔颌面外科,江苏南京210008

出　　处：《陕西医学杂志》2024年第2期157-162,共6页Shaanxi Medical Journal

基　　金：江苏省重点研发计划(社会发展)专项资金资助项目(BE2021609)。

摘　　要：目的:探讨微小核糖核酸-21(miR-21)介导Smad7调节转化生长因子β(TGF-β)/Smad信号通路对人口腔癌细胞株HSQ-89增殖、侵袭与迁移的作用。方法:取人口腔癌细胞株HSQ-89培养传代,分为阴性对照组、下调组、上调组和正常组,前三者分别采用脂质体转染法将携带阴性对照(NC inhibitor)、miR-21抑制剂(miR-21 inhibitor)、miR-21模拟物(miR-21 mimics)载体进行转染,正常组仅添加等量无菌蒸馏水。观察各组细胞增殖、侵袭、迁移能力;实时-定量聚合酶链反应(RT-qPCR)检测各组细胞miR-21、TGF-β_(1)、Smad3、Smad7、细胞周期蛋白D1(CCND1)、MYC、E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)、基质金属蛋白酶-2(MMP-2)mRNA表达;蛋白免疫印迹法(WB)检测各组细胞TGF-β_(1)、Smad3、Smad7、CCND1、MYC、E-cadherin、Vimentin、MMP-2蛋白表达及p-Smad3水平;双荧光素酶报告基因检测验证miR-21是否靶向Smad7。结果:与正常组和阴性对照组比较,上调组细胞增殖、侵袭、迁移能力上升,下调组细胞增殖活性下降、侵袭细胞数减少、划痕愈合率下降(均P<0.05);与正常组和阴性对照组比较,上调组细胞miR-21表达、TGF-β_(1)、Smad3、CCND1、MYC、Vimentin、MMP-2 mRNA和蛋白表达上升、Smad7和E-cadherin mRNA和表达下降,差异有统计学意义(均P<0.05),下调组细胞miR-21表达、TGF-β_(1)、Smad3、CCND1、MYC、Vimentin、MMP-2 mRNA和蛋白表达下降、Smad7和E-cadherin mRNA和蛋白表达上升(均P<0.05);经双荧光素酶报告基因实验验证miR-21靶向Smad7。结论:miR-21促进口腔癌细胞增殖、侵袭和迁移,可能与调节Smad7、TGF-β/Smad信号通路相关。Objective:To investigate the effect of miR-21 mediating Smad7 to regulate transforming growth factorβ(TGF-β)/Smad signaling pathway on the proliferation,invasion and migration of human oral cancer cell lines HSQ-89.Methods:Human oral cancer cell lines HSQ-89 were cultured and subcultured,which was divided into negative control group,down-regulated group,up-regulated group and normal group,The former three were transfected vectors carrying negative control(NC inhibitor),miR-21 inhibitor and miR-21 mimics by liposome transfection method respectively,while the normal group was only added the same amount of sterile distilled water.The cell proliferation,invasive and migration activities were detected.RT-qPCR was used to detect miR-21,TGF-β_(1),Smad3,Smad7,cyclin D1(CCND1),MYC,E-cadherin,Vimentin and matrix metalloproteinase-2(MMP-2)mRNA expressions in each group.The protein expressions of TGF-β_(1),Smad3,Smad7,CCND1,MYC,E-cadherin,Vimentin,MMP-2 and p-Smad3 were detected by Western blot(WB).Dual luciferase reporter gene assay was used to verify whether miR-21 targeted Smad7.Results:Compared with the normal group and the negative control group,the cell proliferation,invasion and migration ability of the up-regulated group were increased,and the cell proliferation activity,the number of invasive cells and the scratch healing rate of the down-regulated group were decreased(all P<0.05).Compared with normal group and negative control group,miR-21 expression,mRNA and protein expressions of TGF-β_(1),Smad3,CCND1,MYC,Vimentin,MMP-2 were increased,while the mRNA and protein expressions of Smad7 and E-cadherin were decreased in up-regulated group(all P<0.05);miR-21 expression,mRNA and protein expressions of TGF-β_(1),Smad3,CCND1,MYC,Vimentin,MMP-2 were decreased,while the mRNA and protein expressions of Smad7 and E-cadherin were increased in down regulated group(all P<0.05).Dual luciferase reporter gene assay confirmed that miR-21 targeted Smad7.Conclusion:MiR-21 promotes the proliferation,invasion and migration of oral

关 键 词：口腔癌 微小核糖核酸-21 转化生长因子Β 细胞增殖 细胞侵袭 细胞迁移 

分 类 号：R739.8[医药卫生—肿瘤]