Coronin 1c过表达促进宫颈癌细胞系SiHa的上皮间质转换

Coronin 1c over-expression promotes epithelial-mesenchymal transition in cervical cancer cell line SiHa

目的基于Notch1/LOX/Snail1通路探索冠蛋白1c(Coronin 1c)过表达对宫颈癌细胞系SiHa的上皮间质转换(EMT)过程的影响。方法构建针对Coronin 1c基因的RNA干扰重组腺病毒(Ad-Cor-siRNA)及含对照序列的重组腺病毒(Ad-control),并感染SiHa细胞沉默Coronin 1c基因表达;并分组为H8组:H8细胞;空白对照(blank)组:未用任何病毒处理的SiHa细胞;Ad-control组:感染空载体腺病毒Ad-control的SiHa细胞;Ad-Cor-siRNA组:感染重组腺病毒Ad-Cor-siRNA的SiHa细胞。Western blot检测细胞中Coronin 1c、E-cadherin、vimentin、LOX及Snail1蛋白表达;qPCR检测细胞E-cadherin和vimentin mRNA表达;免疫荧光实验检测细胞MMP-2、MMP-9和Notch1蛋白表达。结果与正常宫颈上皮细胞H8相比,SiHa细胞中Coronin 1c、vimentin、MMP-2、MMP-9、Notch1、LOX及Snail1蛋白表达均升高,E-cadherin表达下降。Coronin 1c沉默后,可促使vimentin、MMP-2、MMP-9、Notch1、LOX及Snail1蛋白表达下降,E-cadherin表达升高。结论Coronin 1c蛋白过表达与SiHa细胞EMT密切联系,其能激活Notch1/LOX/Snail1通路,促进SiHa细胞EMT。

Objective To investigate the effect of over-expression of Coronin 1c on the epithelial-mesenchymal transition(EMT)of cervical cancer cell line SiHa based on Notch1/LOX/Snail1 pathway.Methods RNA interference recombinant adenovirus targeting Coronin 1c gene(Ad-Cor-siRNA)and recombinant adenovirus containing control sequence(Ad-control)were constructed,and then infected SiHa cells to silence Coronin 1c gene expression;and grouped into H8 group:H8 cells,blank control group(blank):SiHa cells not treated with any virus,Ad-Control group:SiHa cells infected with empty vector adenovirus,Ad-Cor-siRNA group:SiHa cells infected with recombinant adenovirus Ad-Cor-siRNA;Western blot was used to detect the expression of Coronin 1c,E-cadherin,Vimentin,LOX and Snail1 in each group.The expressions of E-cadherin and Vimentin mRNA were detected by qPCR.The expressions of MMP-2,MMP-9 and Notch1 were detected by immunofluorescence assay.Results Compared with normal cervical epithelial cells H8,the expressions of Coronin 1c,Vimentin,MMP-2,MMP-9,Notch1,LOX and Snail1 proteins in SiHa cells were increased,and the expressions of E-cadherin were decreased.After silencing of Coronin 1c,the expressions of Vimentin,MMP-2,MMP-9,Notch1,LOX and Snail1 proteins were decreased,and the expression of E-cadherin was increased.Conclusions The over-expression of Coronin 1c protein is closely related to the EMT transformation of SiHa cells,which can activate the Notch1/LOX/Snail1 pathway and promote EMT transformation in SiHa cells.