Twist-related protein 1 (TWIST1), a highly conserved basic helix-loop-helix transcription factor, stimulates epithelial-mesenchymal transition (EMT) and plays a crucial role in the regulation of the extracellular matrix (ECM) and cell-cell adhesion. Our aim in this study was to evaluate the functional correlation between TWIST1 and MMP genes in human ESCC cell lines, KYSE-30 and YM-1. To generate recombinant retroviral particles, the Pruf-IRES-GFP-hTWIST1 was co-transfected into HEK293T along with pGP and pMD2. G as well as Pruf-IRES-GFP control plasmid. Stably transduced high-expressing GFP-hTWIST1 and GFP-control KYSE-30 cells were generated. The produced retroviral particles were transduced into the KYSE-30 and YM-1 ESCC cells. Ectopic expression of TWIST1 mRNA and expression of the MMP genes (MMP-2, MMP-3, MMP-7, MMP-9, and MMP-10) were examined by relative comparative real-time PCR. In silico analysis of the MMP markers and their promoter elements was explored. Moreover, the scratch wound assay was used to evaluate the migration of TWIST1-induced cells. TWIST1 level was up-regulated by nearly 5-fold and 7.4-fold in GFP-hTWIST1 KYSE-30 and YM-1 cells compared to GFP control cells, respectively. Interestingly, this enforced expression of TWIST1 subsequently caused significant overexpression of transcripts for selected MMP genes in GFP-hTWIST1 in comparison with GFP control cells in both ESCC cell lines. Also, the scratch assay indicated that TWIST1 expression effectively increased the migration of GFP-TWIST1 KYSE-30 cells against GFP KYSE-30 control cells in vitro. The present findings illuminate that TWIST1 may contribute broadly to ESCC development in concert with up-regulation of MMPs expression and further suggest the potential advantage of exerting TWIST1/MMPs signaling axis as a framework from which to expand our understanding about the mechanisms of ESCC tumorigenesis.

扭曲相关蛋白1（TWIST1），一种高度保守的基本螺旋-环-螺旋转录因子，刺激上皮-间质转化（EMT），并在调节细胞外基质（ECM）和细胞粘附方面起关键作用。我们在这项研究中的目的是评估TWIST1和MMP基因在人类ESCC细胞系KYSE-30和YM-1之间的功能相关性。为了产生重组逆转录病毒颗粒，将Pruf-IRES-GFP-hTWIST1与pGP和pMD2一起共转染到HEK293T中。G以及Pruf-IRES-GFP对照质粒。产生稳定转导的高表达GFP-hTWIST1和GFP对照KYSE-30细胞。产生的逆转录病毒颗粒被转导到KYSE-30和YM-1 ESCC细胞中。TWIST1 mRNA的异位表达和MMP基因（MMP-2，MMP-3，MMP-7，MMP-9，和MMP-10）通过相对比较实时PCR进行检查。在计算机上分析了MMP标记及其启动子元件。此外，使用刮擦试验来评估TWIST1诱导的细胞的迁移。与GFP对照细胞相比，GFP-hTWIST1 KYSE-30和YM-1细胞的TWIST1水平分别上调了近5倍和7.4倍。有趣的是，与两种ESCC细胞系中的GFP对照细胞相比，TWIST1的这种强制表达随后导致GFP-hTWIST1中所选MMP基因的转录物显着过表达。而且，划痕试验表明TWIST1表达在体外有效地增加了GFP-TWIST1 KYSE-30细胞相对于GFP KYSE-30对照细胞的迁移。