Department of Biochemistry, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia.
Department of Pharmacology, College of Pharmacy, Jouf University, Sakaka, Al-Jouf, Saudi Arabia.
Crit Rev Eukaryot Gene Expr. 2021;31(2):89-95. doi: 10.1615/CritRevEukaryotGeneExpr.2021037996.
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.
在各种上皮-间充质转化 (EMT) 相关转录因子 (TF) 中，Snail-1、Snail-2/Slug、Twist 和 ZEB1 的表达水平改变与具有较高转移风险的不同癌症显著相关。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 在定义子宫内膜异位症进展和分化的分子发病机制中的关键作用，并与对照组进行比较。对 75 例不同分期的子宫内膜异位症患者和 50 例对照组患者的外周血样本进行了 Snail-1、Snail-2/Slug、Twist 和 ZEB1 的定性和定量表达分析。提取总 RNA 并转化为 cDNA，用于使用 SYBRGreen 基于逆转录定量聚合酶链反应 (RT-qPCR) 对每个基因转录本进行相对定量。相对定量的 Livak 方法用于计算与内参相比每个 TF 的倍数变化。与对照组相比，所有四种选定的 TF 在子宫内膜异位症患者中均表现出显著上调的表达水平。与子宫内膜异位症患者的 I 期和 II 期相比，Snail-1 增加了三倍 (p = 0.0001)，Snail-2 增加了两倍 (p = 0.01)，Twist 增加了两倍 (p = 0.0002)，ZEB1 增加了两倍 (p = 0.001)。 。本研究表明，EMT 相关 TF 通过使用非侵入性工具对特定分子级联进行表达分析，在子宫内膜异位症患者的发病机制和区分不同阶段中发挥关键作用。
