子宫内膜癌（EC）是常见的妇科癌症。研究表明EC对5-氟尿嘧啶（5-FU）化疗的敏感性降低，导致治疗效果不理想。迫切需要调查5-FU治疗EC效果不佳的原因。本研究的目的是探讨RAD51AP1被E2F7转录激活后通过脂肪酸代谢途径对EC细胞对5-FU化疗敏感性的影响。EC的mRNA表达数据从癌症基因组图谱数据库下载，进行差异表达分析，并根据生物信息学分析和文献查阅确定目的基因。使用 hTFtarget 数据库预测 EC 中 RAD51AP1 上游的调控转录因子。通过qRT-PCR和蛋白质印迹测量E2F7和RAD51AP1的表达。然后，通过染色质免疫沉淀 (ChIP) 和双荧光素酶测定验证了 E2F7 和 RAD51AP1 之间的转录激活关系。采用CCK-8法测定EC细胞对5-FU的IC50值，流式细胞术检测细胞凋亡。 Western blot检测细胞凋亡相关蛋白和脂肪酸代谢相关蛋白的表达情况。生物信息学分析显示EC中E2F7和RAD51AP1均高表达，并预测了RAD51AP1启动子与E2F7可能的结合位点。 ChIP 测定和双荧光素酶测定证实了 E2F7 与 RAD51AP1 启动子区域的结合。细胞实验表明，过表达RAD51AP1可以促进EC细胞的生长和脂肪酸代谢，并抑制细胞对5-FU的敏感性，而沉默E2F7可以降低RAD51AP1过表达对EC细胞生长和对5-FU敏感性的影响。 E2F7/RAD51AP1轴可通过调节脂肪酸代谢促进EC细胞生长并抑制细胞对5-FU的敏感性，提示E2F7/RAD51AP1轴可能是EC治疗的新途径。版权所有©2023国际抗癌研究所（博士） . 乔治·J·德利纳西奥斯 (George J. Delinasios)，保留所有权利。

Endometrial cancer (EC) is a frequent gynecological cancer. Studies have demonstrated that the sensitivity of EC toward 5-fluorouracil (5-FU) chemotherapy has decreased, leading to unsatisfactory treatment effects. There is an urgent need to investigate the reasons for the unsatisfactory treatment of EC with 5-FU. The purpose of the study was to investigate the effect of RAD51AP1 after being transcriptionally activated by E2F7 on the sensitivity of EC cells to 5-FU chemotherapy via the fatty acid metabolic pathway.mRNA expression data on EC were downloaded from The Cancer Genome Atlas database, subjected to differential expression analysis, and the target genes were determined based on the bioinformatics analysis and literature consulting. The regulatory transcription factor upstream of RAD51AP1 in EC was predicted using the hTFtarget database. The expression of E2F7 and RAD51AP1 was measured by qRT-PCR and western blot. Then, the transcriptional activation relationship between E2F7 and RAD51AP1 was verified by chromatin immunoprecipitation (ChIP) and dual luciferase assays. The IC50 values of EC cells toward 5-FU were determined by the CCK-8 assay, and cell apoptosis was detected by flow cytometry. The expression of apoptosis-related and fatty acid metabolism-related proteins was evaluated by western blot.Bioinformatics analysis showed that both E2F7 and RAD51AP1 were highly expressed in EC, and the possible binding sites between RAD51AP1 promoter and E2F7 were predicted. ChIP assay and dual luciferase assay confirmed the binding of E2F7 to RAD51AP1 promoter region. Cell experiments showed that overexpressing RAD51AP1 could facilitate the growth and fatty acid metabolism of EC cells, and suppress cell sensitivity to 5-FU, while silencing of E2F7 could reduce the effect of RAD51AP1 overexpression on EC cell growth and sensitivity toward 5-FU.The E2F7/RAD51AP1 axis can promote the growth of EC cells and inhibit cell sensitivity to 5-FU by regulating fatty acid metabolism, suggesting that E2F7/RAD51AP1 axis may be a novel pathway for EC treatment.Copyright © 2023 International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.