Wang Yong, Tan Yudi, Yang Shasha, Wei Jinkong, Wei Yuying, Chen Junying
Department of Gynecology, The First Affiliated Hospital of Guangxi Medical University, Nanning, China.
Gynecol Obstet Invest. 2025;90(2):108-119. doi: 10.1159/000540384. Epub 2024 Sep 30.
Cervical carcinoma (CC) is prevalent among women worldwide with increasing risk. Finding effective methods for treating CC is of utmost importance. The aim of this study was to investigate the effect of SERPINE1 on the progression of cervical precancerous lesions to CC.
This study used transcriptome sequencing and in vitro cell line. Participants/Materials: Cervical precancerous lesions and CC samples and human cervical epithelial immortalized cell line H8, human CC cell lines HeLa, and CaSki were involved in this study.
Next-generation sequencing was applied to identify 100 differentially expressed genes from cervical precancerous lesions and CC samples. With the application of the Search Tool for the Retrieval of Interacting Genes (STRING) database, we carried out the protein-protein interaction network analysis, thus screening out serine protease inhibitor clade E member 1 (SERPINE1) with significant upregulation in CC cells. The helicase-like transcription factor (HLTF) was predicted as the upstream transcription factor using Human Transcription Factor Database (HumanTFDB). The chromatin immunoprecipitation (ChIP) experiment was conducted to validate the interaction between SERPINE1 and HLTF. The immunohistochemistry was employed to determine the expression of SERPINE1 and HLTF in CC tissues. Following the upregulation or downregulation of SERPINE1 and HLTF, the real-time quantitative reverse transcription polymerase chain reaction was carried out to assess mRNA expression levels of SERPINE1 and HLTF in cells. Cell viability, migration, and invasion were evaluated using MTT assay, cell scratch assay, and Transwell assay, respectively. Western blot analysis was conducted to assess changes in the expression levels of matrix metalloproteinases and proteins related to epithelial-mesenchymal transition (EMT).
The ChIP experiment confirmed the interaction between HLTF and SERPINE1. HLTF and SERPINE1 were upregulated in CC tissues and cells, and silencing SERPINE1 inhibited the EMT process and viability, migration, and invasion of CC cells. However, overexpression of SERPINE1 in CC cells showed the opposite trend. Rescue experiments demonstrated that silencing HLTF repressed CC cell viability, migration, and invasion, which could be restored by overexpressing SERPINE1.
The effect of the HLTF/SERPINE1 axis on CC malignant progression has not been confirmed by in vivo experiments.
HLTF transcriptionally activates SERPINE1, promoting the progression from cervical precancerous lesions to CC.
宫颈癌（CC）在全球女性中普遍存在且风险不断增加。寻找有效的CC治疗方法至关重要。本研究旨在探讨丝氨酸蛋白酶抑制剂E1家族成员1（SERPINE1）对宫颈上皮内瘤变进展为CC的影响。
本研究采用转录组测序和体外细胞系。参与者/材料：本研究纳入了宫颈上皮内瘤变和CC样本以及人宫颈上皮永生化细胞系H8、人CC细胞系HeLa和CaSki。
应用二代测序从宫颈上皮内瘤变和CC样本中鉴定出100个差异表达基因。利用检索相互作用基因的搜索工具（STRING）数据库进行蛋白质-蛋白质相互作用网络分析，从而筛选出在CC细胞中显著上调的丝氨酸蛋白酶抑制剂E1家族成员1（SERPINE1）。使用人类转录因子数据库（HumanTFDB）预测解旋酶样转录因子（HLTF）为上游转录因子。进行染色质免疫沉淀（ChIP）实验以验证SERPINE1与HLTF之间的相互作用。采用免疫组织化学法测定CC组织中SERPINE1和HLTF的表达。在SERPINE1和HLTF上调或下调后，进行实时定量逆转录聚合酶链反应以评估细胞中SERPINE1和HLTF的mRNA表达水平。分别使用MTT法、细胞划痕实验和Transwell实验评估细胞活力、迁移和侵袭能力。进行蛋白质印迹分析以评估基质金属蛋白酶和上皮-间质转化（EMT）相关蛋白表达水平的变化。
ChIP实验证实了HLTF与SERPINE1之间的相互作用。HLTF和SERPINE1在CC组织和细胞中上调，沉默SERPINE1可抑制CC细胞的EMT过程以及活力、迁移和侵袭能力。然而，CC细胞中SERPINE1的过表达呈现相反趋势。挽救实验表明，沉默HLTF可抑制CC细胞活力、迁移和侵袭能力，而过表达SERPINE1可使其恢复。
HLTF/SERPINE1轴对CC恶性进展的影响尚未通过体内实验得到证实。
HLTF转录激活SERPINE1，促进宫颈上皮内瘤变进展为CC。
