调控叉头盒转录基因F1表达对非小细胞肺癌细胞上皮间质转化、迁移和侵袭的作用

Effect of forkhead box-F1 expression on epithelial-mesenchymal transition,migration and invasion of nonsmall cell lung cancer cells and its mechanism

目的通过慢病毒介导的shRNA敲低人非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞株PC9中叉头盒转录基因F1(forkhead box-F1,FOXF1)的表达,研究FOXF1对细胞生长及侵袭迁移等能力的影响及可能的分子机制。方法慢病毒介导的shRNA敲低FOXF1的表达,采用实时荧光定量qRTPCR实验及Western Blot技术检测细胞中FOXF1 mRNA和蛋白的表达变化,平板克隆实验及CCK-8法检测细胞增殖情况,Transwell实验检测FOXF1基因表达对迁移及侵袭的影响,Western Blot检测上皮间充质转化相关蛋白的表达。结果慢病毒介导的shRNA感染PC9细胞后,有效下调了PC9细胞中FOXF1 mRNA和蛋白的表达水平(P <0.05),细胞的增殖能力受到抑制,细胞体外迁移及侵袭能力明显降低;Western Blot结果显示,抑制FOXF1表达后β-Catenin、N-cadherin、Vimentin、Snail表达下调,E-cadherin、ZO-1表达上调。结论下调FOXF1表达可抑制非小细胞肺癌PC9细胞的迁移和侵袭能力,其机制可能通过调控上皮—间充质转化相关蛋白的表达来实现。

Objective Through knock down the expression of Forkhead box-F1(FOXF1)in human non-small cell lung cancer(NSCLC)cell line PC9 by lentiviral-mediated sh RNA,to explore the effects of FOXF1 inproliferation,epithelia-mesenchymal transition,migration and invasion and its related-mechanism.Methods Lentivirus-mediated sh RNA knocked down FOXF1 expression in PC9 cells,and the expression of FOXF1 m RNAand protein in infected cells were detected by q RT-PCR and Western Blot. The cell proliferation was detected byColony-forming assay and CCK-8 assay. The invasion and migration ability of cells were detected by Transwellassay. Western Blot was used to detect the expression of proteins related to epithelia-mesenchymal transition.ResultsLentiviral-mediated sh RNA significantly enhanced the protein and m RNA expression of FOXF1 in PC9 cells(P < 0.05). Cell proliferation was inhibited,and cell migration and invasion in vitro were decreased. WesternBlot results showed the expression of β-Catenin,N-cadherin,Vimentin,and Snail were down-regulated,but theexpression of E-cadherin and ZO-1 were up-regulated.ConclusionKnocking down FOXF1 expression can inhibitthe migration and invasion ability of non-small cell lung cancer cell line PC9 cells,which may be achieved byregulating the expression of proteins related to epithelial-mesenchymal transition.