Abstract：
The purpose of this RNA sequencing study was to investigate the biological mechanism underlying how the transcription factors (TFs) Twist1 and Zeb1 influence the prognosis of mycosis fungoides (MF). We used laser-captured microdissection to dissect malignant T-cells obtained from 40 skin biopsies from 40 MF patients with stage I-IV disease. Immunohistochemistry (IHC) was used to determinate the protein expression levels of Twist1 and Zeb1. Based on RNA sequencing, principal component analysis (PCA), differential expression (DE) analysis, ingenuity pathway analysis (IPA), and hub gene analysis were performed between the high and low Twist1 IHC expression cases. The DNA from 28 samples was used to analyze the TWIST1 promoter methylation level. In the PCA, Twist1 IHC expression seemed to classify cases into different groups. The DE analysis yielded 321 significant genes. In the IPA, 228 significant upstream regulators and 177 significant master regulators/causal networks were identified. In the hub gene analysis, 28 hub genes were found. The methylation level of TWIST1 promoter regions did not correlate with Twist1 protein expression. Zeb1 protein expression did not show any major correlation with global RNA expression in the PCA. Many of the observed genes and pathways associated with high Twist1 expression are known to be involved in immunoregulation, lymphocyte differentiation, and aggressive tumor biology. In conclusion, Twist1 might be an important regulator in the disease progression of MF.
摘要：
这项RNA测序研究的目的是研究转录因子（TFs）Twist1和Zeb1如何影响真菌病（MF）预后的生物学机制。我们使用激光捕获的显微解剖法来解剖从40例I-IV期MF患者的40例皮肤活检中获得的恶性T细胞。免疫组织化学(IHC)用于测定Twist1和Zeb1的蛋白表达水平。基于RNA测序，主成分分析(PCA)，差异表达（DE）分析，创新途径分析(IPA)，在高和低Twist1IHC表达病例之间进行了hub基因分析。来自28个样品的DNA用于分析TWISTl启动子甲基化水平。在PCA中,Twist1IHC表达似乎将病例分为不同的组。DE分析产生321个重要基因。在IPA中,确定了228个重要的上游监管机构和177个重要的主监管机构/因果网络。在枢纽基因分析中，发现了28个hub基因。TWIST1启动子区的甲基化水平与Twist1蛋白表达无关。在PCA中，Zeb1蛋白表达与全局RNA表达没有任何主要相关性。已知许多观察到的与Twist1高表达相关的基因和途径参与免疫调节,淋巴细胞分化，和侵袭性肿瘤生物学。总之,Twist1可能是MF疾病进展的重要调节因子。
