Chen Hua, Han Cailing, Ha Chunfang
Department of Gynecology, General Hospital of Ningxia Medical University, Shengli South Street, Xingqing District, Yinchuan, 750004, Ningxia, China.
Department of Anesthesiology, General Hospital of Ningxia Medical University, Yinchuan, 750004, Ningxia, China.
Appl Biochem Biotechnol. 2025 Apr;197(4):2133-2150. doi: 10.1007/s12010-024-05116-w. Epub 2024 Dec 14.
Uterine corpus endometrial carcinoma (UCEC) is one of the most common gynecologic tumors. Due to the high recurrence and metastasis of UCEC, it is crucial for patients to find new biomarkers for diagnosis and therapy. In this study, R software and the TCGA database were used to screen candidate UCEC predictive markers. Western blot and RT-qPCR were performed to detect protein and mRNA expression of EXT1 in UCEC cell lines. In addition, MTT assay, flow cytometry, transwell assay, and wound healing assay were conducted to assess the cell viability, apoptosis, invasion, and migration in UCEC cells. Overlap-extension PCR technique was employed to construct the vector targeting the deletion of the methylated segment of EXT1. The results showed that a total of 11 candidate genes were obtained and EXT1 was identified as a potential target. The expression and methylation levels of EXT1 were both increased in UCEC tissues and cell lines, as well as elevated EXT1 was closely related to the poor prognosis of patients. Besides, the knockdown of EXT1 significantly inhibited the malignant biological behaviors in UCEC cells. Additionally, the current study also found that the deletion of 1559-2146 bp CpG island segment upregulated EXT1 expression and promoted malignant biological behaviors in UCEC cells. Furthermore, the presence of m7G RNA methylation in UCEC cells also was found. In conclusion, the methylation of EXT1 influenced the gene expression, thereby affecting the malignant biological behaviors in UCEC cells and regulating the pathological progression of UCEC.
子宫体子宫内膜癌（UCEC）是最常见的妇科肿瘤之一。由于UCEC的高复发和转移率，寻找新的诊断和治疗生物标志物对患者至关重要。在本研究中，使用R软件和TCGA数据库筛选UCEC候选预测标志物。采用蛋白质免疫印迹法和逆转录定量聚合酶链反应（RT-qPCR）检测UCEC细胞系中EXT1的蛋白质和mRNA表达。此外，进行MTT法、流式细胞术、Transwell实验和伤口愈合实验，以评估UCEC细胞的活力、凋亡、侵袭和迁移能力。采用重叠延伸PCR技术构建靶向EXT1甲基化片段缺失的载体。结果显示，共获得11个候选基因，EXT1被确定为潜在靶点。EXT1在UCEC组织和细胞系中的表达及甲基化水平均升高，且EXT1升高与患者预后不良密切相关。此外，敲低EXT1可显著抑制UCEC细胞的恶性生物学行为。另外，本研究还发现，缺失1559-2146 bp CpG岛片段可上调UCEC细胞中EXT1的表达并促进其恶性生物学行为。此外，还发现UCEC细胞中存在m7G RNA甲基化。总之，EXT1的甲基化影响基因表达，从而影响UCEC细胞的恶性生物学行为并调节UCEC的病理进展。
