Wang Jing, Cong Shanshan, Wu Han, He Yanan, Liu Xiaoli, Sun Liyuan, Zhao Xibo, Zhang Guangmei
Department of Gynecology, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
Department of Gynecology, The Red Cross Center Hospital of Harbin, Harbin, China.
Front Mol Biosci. 2021 Nov 1;8:743012. doi: 10.3389/fmolb.2021.743012. eCollection 2021.
Endometriosis is a serious gynecological disorder characterized by debilitating pain, infertility and the establishment of innervated endometriosis lesions outside the uterus. Early detection and accurate diagnosis are pivotal in endometriosis. The work screened autophagy-related genes (ATGs) as potential biomarkers to reveal new molecular subgroups for the early diagnosis of endometriosis. The gene lists of ATGs from five databases were integrated. Then, weighted gene co-expression network analysis (WGCNA) was used to map the genes to the gene profile of endometriosis samples in GSE51981 to obtain functional modules. GO and KEGG analyses were performed on the ATGs from the key modules. Differentially expressed ATGs were identified by the limma R package and further validated in the external datasets of GSE7305 and GSE135485. The DESeq2 R package was utilized to establish multifactorial network. Subsequently, one-way analysis of variance (ANOVA) was performed to identify new molecular subgroups. Real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting were used to confirm the differential expression of hub ATGs, and the receiver operating characteristic (ROC) curve analysis and Spearman correlation analysis were applied to assess the diagnostic value of hub ATGs in 40 clinical samples and human primary endometrial stromal cells (ESCs). We screened 4 key modules and 12 hub ATGs and found the key genes to be strongly correlated with endometriosis. The pathways of ATGs were mainly enriched in autophagy, apoptosis, ubiquitin-protein ligase binding, and MAPK signaling pathway. The expression levels of EZH2 (Enhancer of Zeste homolog 2) and RND3 (also known as RhoE) had statistically significant changes with higher values in the endometriosis group compared with the controls, both in the tissue samples and primary ESCs. Besides, they also showed higher specificity and sensitivity by the receiver operating characteristic analysis and Spearman correlation analysis for the diagnosis of endometriosis. The TF-mRNA-miRNA-lncRNA multifactorial network was successfully constructed. Four new molecular subgroups were identified, and we preliminarily showed the ability of IQCG to independently differentiate subgroups. EZH2 and RND3 could be candidate biomarkers for endometriosis, which would contribute to the early diagnosis and intervention in endometriosis.
子宫内膜异位症是一种严重的妇科疾病，其特征为使人衰弱的疼痛、不孕以及子宫外出现有神经支配的子宫内膜异位病变。早期检测和准确诊断在子宫内膜异位症中至关重要。这项研究筛选了自噬相关基因（ATG）作为潜在生物标志物，以揭示用于子宫内膜异位症早期诊断的新分子亚组。整合了来自五个数据库的ATG基因列表。然后，使用加权基因共表达网络分析（WGCNA）将这些基因映射到GSE51981中子宫内膜异位症样本的基因谱上，以获得功能模块。对关键模块中的ATG进行了基因本体（GO）和京都基因与基因组百科全书（KEGG）分析。通过limma R包鉴定差异表达的ATG，并在GSE7305和GSE135485的外部数据集中进一步验证。利用DESeq2 R包建立多因素网络。随后，进行单因素方差分析（ANOVA）以识别新的分子亚组。采用实时定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法来确认核心ATG的差异表达，并应用受试者工作特征（ROC）曲线分析和Spearman相关性分析来评估核心ATG在40例临床样本和人原代子宫内膜基质细胞（ESC）中的诊断价值。我们筛选出4个关键模块和12个核心ATG，发现这些关键基因与子宫内膜异位症密切相关。ATG的途径主要富集在自噬、凋亡、泛素-蛋白连接酶结合和丝裂原活化蛋白激酶（MAPK）信号通路中。在组织样本和原代ESC中，与对照组相比，子宫内膜异位症组中zeste同源物2增强子（EZH2）和RND3（也称为RhoE）的表达水平有统计学意义的变化，且值更高。此外，通过ROC分析和Spearman相关性分析，它们在诊断子宫内膜异位症时也表现出更高的特异性和敏感性。成功构建了转录因子-信使核糖核酸-微小核糖核酸-长链非编码核糖核酸多因素网络。识别出四个新的分子亚组，并且我们初步展示了IQCG独立区分亚组的能力。EZH2和RND3可能是子宫内膜异位症的候选生物标志物，这将有助于子宫内膜异位症的早期诊断和干预。
