Among various epithelial-to-mesenchymal transition (EMT)-related transcription factors (TFs), altered expression levels of Snail-1, Snail-2/Slug, Twist, and ZEB1 have shown a significant association in different cancers having a higher risk of metastasis. However, their role in the circulation of endometriosis patients is not well understood. Hence, the present study was designed to evaluate the crucial role of these TFs in defining the molecular pathogenesis for endometriosis progression and differentiation from control subjects. The qualitative and quantitative expression analysis of Snail-1, Snail-2/Slug, Twist, and ZEB1 were analyzed in peripheral blood samples of 75 different stages of endometriosis patients and compared with 50 control subjects. Total RNA was extracted and converted into complementary DNA (cDNA) for relative quantification of each gene transcript using SYBRGreen-based reverse transcriptase quantitative polymerase chain reaction (RT-qPCR). The Livak method of relative quantification was used for calculating the fold change in each TF compared with endogenous control. All four selected TFs showed significantly upregulated expression levels in endometriosis patients compared with control subjects. A three-fold increase was observed for Snail-1 (p = 0.0001), and a two-fold increase was observed for Snail-2 (p = 0.01), Twist (p = 0.0002), and ZEB1 (p = 0.001) in stage III and IV compared with stage I and II of endometriosis patients. The present study revealed that EMT-related TFs play a crucial role in the pathogenesis and differentiating different stages of endometriosis patients through expression analysis of specific molecular cascades using non-invasive tools.

中文翻译：

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上皮-间质转化标志物在子宫内膜异位症不同阶段的作用：Snail、Slug、ZEB1 和 Twist 基因的表达。

在各种上皮-间质转化 (EMT) 相关转录因子 (TF) 中，Snail-1、Snail-2/Slug、Twist 和 ZEB1 的表达水平改变已显示出与具有较高风险的不同癌症的显着关联转移。然而，它们在子宫内膜异位症患者循环中的作用尚不清楚。因此，本研究旨在评估这些 TF 在确定子宫内膜异位症进展的分子发病机制和与对照受试者分化中的关键作用。对 75 名不同阶段子宫内膜异位症患者的外周血样本进行 Snail-1、Snail-2/Slug、Twist 和 ZEB1 的定性和定量表达分析，并与 50 名对照受试者进行比较。使用基于 SYBRGreen 的逆转录酶定量聚合酶链式反应 (RT-qPCR) 提取总 RNA 并将其转化为互补 DNA (cDNA) 以对每个基因转录物进行相对定量。相对定量的 Livak 方法用于计算每个 TF 与内源对照相比的倍数变化。与对照受试者相比，所有四种选定的 TF 在子宫内膜异位症患者中均显示出显着上调的表达水平。Snail-1 (p = 0.0001) 增加了三倍，Snail-2 (p = 0.01)、Twist (p = 0.0002) 和 ZEB1 (p = 0.001) 增加了两倍III 期和 IV 期子宫内膜异位症患者与 I 期和 II 期相比。