Aberrant expression of cancer testis antigens (CTAs) is reported in tumors, especially those with stemness properties. A number of CTAs can induce epithelial mesenchymal transition (EMT) process and promote cancer stem cells (CSCs) characteristics. We aimed in this study to analyze the correlation between NY-ESO1 and TWIST1 in esophageal squamous cell carcinoma (ESCC), as well as their impact on EMT process. Gene expression profiling of NY-ESO1 and TWIST1 was performed in 43 esophageal tumors compared to their margin normal tissues of using qRT-PCR, and their correlation with clinicopathological variables of the patients was evaluated. In silico analysis of the NY-ESO1, epithelial and mesenchymal cell markers and also their promoter sequences was executed. ESCC cell lines KYSE-30 and YM-1 were transduced to ectopically express TWIST1 using a retroviral system, followed by qRT-PCR mRNA expression analysis to reveal the probable correlation among TWIST1, NY-ESO1 and EMT markers gene expression. Scratch assay was performed to estimate migration of TWIST1-induced cells. Overexpression of TWIST1 and NY-ESO1 mRNA was observed in 42% and 39.5% (P ˂ 0.05) of tumors, respectively. Expression of the genes was significantly correlated with each other (p = 0.005). TWIST1 and NY-ESO1 overexpression was significantly associated with stage of progression and size of tumors, respectively. A direct association between TWIST1 and NY-ESO1 mRNA expression was confirmed by induced ectopic expression of TWIST1 in ESCC cell lines KYSE-30 and YM-1. TWIST1-induced cells led to increase migration in ESCC cell line.

Furthermore, significant up-regulation of EMT markers was observed following ectopic expression of TWIST1 in these cells. Based on our findings, it may be proposed that a vital association is exist between the EMT and the acquisition of cancer stemness state in tumor cells through the TWIST1/NY-ESO1 axis and it can be a critical hallmark in ESCC tumorigenesis.

据报道，在肿瘤中，尤其是具有干性的肿瘤中，睾丸抗原（CTA）的异常表达。许多CTA可以诱导上皮间质转化（EMT）过程并促进癌症干细胞（CSCs）特性。在这项研究中，我们旨在分析食管鳞状细胞癌（ESCC）中NY-ESO1和TWIST1之间的相关性，以及它们对EMT过程的影响。使用qRT-PCR与43例正常边缘组织相比，在43例食管肿瘤中进行了NY-ESO1和TWIST1的基因表达谱分析，并评估了它们与患者临床病理变量的相关性。在对NY-ESO1的计算机分析中，执行了上皮和间充质细胞标记物以及它们的启动子序列。使用逆转录病毒系统转导ESCC细胞系KYSE-30和YM-1异位表达TWIST1，然后进行qRT-PCR mRNA表达分析，以揭示TWIST1，NY-ESO1和EMT标记物基因表达之间可能的相关性。进行刮擦测定以估计TWIST1诱导的细胞的迁移。TWIST1和NY-ESO1 mRNA的过表达分别在42％和39.5％（P observed 0.05）的肿瘤中观察到。基因的表达彼此之间显着相关（p = 0.005）。TWIST1和NY-ESO1的过表达分别与肿瘤的进展阶段和大小显着相关。通过在ESCC细胞系KYSE-30和YM-1中诱导的TWIST1异位表达，证实了TWIST1和NY-ESO1 mRNA表达之间的直接关联。TWIST1诱导的细胞导致ESCC细胞系迁移增加。

此外，在这些细胞中异位表达TWIST1后，观察到EMT标记显着上调。根据我们的发现，可能建议在EMT与通过TWIST1 / NY-ESO1轴获取肿瘤细胞中的癌干状态之间存在重要的联系，这可能是ESCC肿瘤发生中的关键标志。