SIX1和TGF-β对原代人喉鳞癌细胞上皮—间质转化的影响  

Effect of SIX1 and TGF-βon epithelial-mesenchymal transition in the human laryngeal squamous cell of primary cultured

作　　者：张继华[1] 董凯峰[1] 苏静[1] 李丹[1] 田君海[1] Zhang Jihua;Dong Kaifeng;Su Jing;Li Dan;Tian Junhai(Department of Otolaryngology, The First Hospital of Hebei Medical University, Hebei Province, Shijiazhuang 050031, China)

机构地区：[1]河北医科大学第一医院耳鼻咽喉科,石家庄050031

出　　处：《疑难病杂志》2020年第4期389-393,398,共6页Chinese Journal of Difficult and Complicated Cases

基　　金：河北省科学技术计划项目(162777262)。

摘　　要：目的探讨原代培养的人喉鳞癌细胞中SIX1和转化生长因子-β(TGF-β)共同作用对上皮—间质转化(EMT)的影响及相关机制。方法收集2016年1月—2018年1月于河北医科大学第一医院耳鼻咽喉科行喉鳞癌手术患者21例,均留存切除组织,并对其进行人喉鳞癌原代细胞培养,制备SIX1、TGF-β及SIX1+TGF-β靶向siRNA,并转染至原代人喉鳞癌细胞,成功转染后分5组,即未转染组、空转组、SIX1转染组、TGF-β转染组及SIX1+TGF-β转染组。应用免疫组化荧光法及蛋白印记Western-blot法检测细胞中SIX1、TGF-β及上皮标志物E-cadherin的蛋白表达。PCR方法检测各组细胞中SIX1、TGF-β及E-cadherin mRNA表达。结果与未转染组比较,空转组细胞中SIX1、TGF-β、E-cadherin mRNA及蛋白表达比较差异均无统计学意义(P>0.05),SIX1转染组、TGF-β转染组及SIX1+TGF-β转染组E-cadherin蛋白及mRNA表达均上调,差异均有统计学意义(F/P=6.846/0.006、13.396/0.000、9.673/0.000)。SIX1转染组、TGF-β转染组及SIX1+TGF-β转染组3组比较,E-cadherin mRNA及其蛋白表达量差异无统计学意义(P>0.05)。结论人喉鳞癌细胞上皮—间质转化呈SIX1及TGF-β依赖性。SIX1和TGF-β共同作用可能是人喉鳞癌细胞上皮—间质转化的关键;SIX1和TGF-β是人喉鳞癌细胞转移潜能的重要标志。Objective To investigate the effects of SIX1 and TGF-βon epithelial-mesenchymal transition(EMT)and related mechanisms in the human laryngeal squamous cell of Primary culture.Methods 21 patients operated at the First Hospital of Hebei Medical University from January 2016 to January 2018.The human laryngeal squamous cell carcinoma tissue was preserved during surgery.Primary laryngeal squamous carcinoma cell culture was performed in vitro.Preparation SIX1-siRNA,TGF-β-siRNA for transfecting into laryngeal squamous cell carcinoma,after successfully transfected,cells were divided into five groups,un-transfected,empty vector,SIX1 transfected,TGF-βtransfected and SIX1+TGF-βtransfected.The protein expression of SIX1,TGF-β,and E cadherin was determined by Western blotting and immunohistochemistry.The mRNA of SIX1,TGF-β,and E cadherin was determined by PCR.Results Compared with the un-transfected group,all of SIX1,TGF-β,E cadherin protein and mRNA expression in empty vector group had no difference(P>0.05),and the protein and mRNA expression in SIX1 transfected,TGF-βtransfected and SIX1+TGF-βtransfected group all show an up-regulated(F/P=6.846/0.006,F/P=13.396/0.000).Among SIX1 transfected,TGF-βtransfected and SIX1+TGF-βtransfected group,the expression of E cadherin mRNA and protein had no obvious change,the difference was not statistically significant(P>0.05).Conclusion Laryngeal squamous cell carcinoma EMT is SIX1 and TGF-βdependent.The interaction of SIX1 with TGF-βmay be the key to EMT in human laryngeal squamous cell carcinoma cells.SIX1 and TGF-βare an important marker of the metastatic potential of human laryngeal squamous cell carcinoma.

关 键 词：上皮—间质转化 SIX1 转化生长因子-Β 人喉鳞癌细胞 

分 类 号：R73[医药卫生—肿瘤]