Liu Junru, Lu Caijuan
Department of Endocrinology, Jinhua People's Hospital, No. 267 Danxi East Road, Jinhua, 321000, Zhejiang Province, China.
Department of Obstetrics, Hangzhou First People's Hospital, No. 261 Huansha Road, Hangzhou, 310006, Zhejiang Province, China.
Biochem Genet. 2025 Apr;63(2):1381-1399. doi: 10.1007/s10528-024-10769-7. Epub 2024 Apr 2.
Gestational diabetes mellitus (GDM) is one of the most common metabolic diseases in pregnant women, posing significant risks to the life and health of both mothers and fetuses. With improving living standards, the incidence of GDM is increasing rapidly. Therefore, understanding the underlying mechanism of GDM is of paramount importance. We downloaded two datasets from the Gene Expression Omnibus (GEO) database, containing sequencing data specifically related to "gestational diabetes" and "placenta". By merging these two datasets, a mRNA expression dataset was obtained and subjected to bioinformatics analyses. To screen out corresponding genes, differential analysis and weighted correlation network analysis (WGCNA) were carried out. Lasso, support vector machine and random forest analyses were subsequently performed for identifying key genes from the differentially expressed genes (DEGs) jointly screened out through differential analysis and WGCNA. Afterwards, immunoinfiltration and correlation analysis were performed to screen immune cells that play a role in disease progression and explore the correlation between the screened key genes and immune cells, after which Western Blot, quantitative real-time polymerase chain reaction, Immunohistochemistry, methyl thiazolyl tetrazolium, flow cytometry, scratch and Transwell assays were, respectively, performed for verification. For further verification, we found that the expression levels of MAP6D1 and SCUBE1 in embryonic tissues of GDM patients was higher compared to those of healthy pregnant women, which was consistent with the results of bioinformatics analysis. Consequently, SCUBE1 was selected for follow-up experiment. In order to explore the role of SCUBE1 in the development of GDM, we treated the trophoblastic cells HTR-8/SVneo with high glucose, and on this basis downregulated the expression of SCUBE1. Through further analysis, we observed that SCUBE1 had a role in reducing cell activity, migration and invasion, and promoting cell apoptosis. In summary, SCUBE1 promotes the development of GDM by increasing cell apoptosis and reducing cell activity, migration, and invasion.
妊娠期糖尿病（GDM）是孕妇中最常见的代谢性疾病之一，对母亲和胎儿的生命健康构成重大风险。随着生活水平的提高，GDM的发病率正在迅速上升。因此，了解GDM的潜在机制至关重要。我们从基因表达综合数据库（GEO）下载了两个数据集，其中包含与“妊娠期糖尿病”和“胎盘”特别相关的测序数据。通过合并这两个数据集，获得了一个mRNA表达数据集并进行了生物信息学分析。为了筛选出相应的基因，进行了差异分析和加权基因共表达网络分析（WGCNA）。随后进行套索分析、支持向量机分析和随机森林分析，以从通过差异分析和WGCNA共同筛选出的差异表达基因（DEG）中识别关键基因。之后，进行免疫浸润和相关性分析，以筛选出在疾病进展中起作用的免疫细胞，并探索筛选出的关键基因与免疫细胞之间的相关性，随后分别进行蛋白质免疫印迹、定量实时聚合酶链反应、免疫组织化学、噻唑蓝、流式细胞术、划痕试验和Transwell试验进行验证。为了进一步验证，我们发现GDM患者胚胎组织中MAP6D1和SCUBE1的表达水平高于健康孕妇，这与生物信息学分析结果一致。因此，选择SCUBE1进行后续实验。为了探讨SCUBE1在GDM发生发展中的作用，我们用高糖处理滋养层细胞HTR-8/SVneo，并在此基础上下调SCUBE1的表达。通过进一步分析，我们观察到SCUBE1具有降低细胞活性、迁移和侵袭以及促进细胞凋亡的作用。总之，SCUBE1通过增加细胞凋亡以及降低细胞活性、迁移和侵袭来促进GDM的发展。
