Xiao Xin, Huang Tingyun, Chen Bin, Zhu Jinshu, Xiao Qingbang, Bao Yuxin
Institute of Life Sciences, Zunyi Medical University, Zunyi, Guizhou, China.
Key Laboratory of Oral Disease of Higher Schools in Guizhou Province, Zunyi, Guizhou, China.
Histol Histopathol. 2025 Apr;40(4):509-521. doi: 10.14670/HH-18-793. Epub 2024 Jul 11.
Endometrial cancer (EC) remains a prevalent gynecological disease with a continuously rising incidence and fatality rate. Acyl coenzyme A: cholesterol acyltransferase 2 (ACAT2) has been commonly perceived as a tumor promoter in multiple human malignancies. This study was conducted to specify the role and mechanism of ACAT2 in EC, which has not been covered. The expression and prognostic significance of ACAT2 in EC samples were respectively analyzed by the ENCORI and Kaplan-Meier plotter databases. RT-qPCR and western blot examined ACAT2 and forkhead box protein A2 (FOXA2) expression in EC cells. The CCK-8 method, colony formation, and EdU staining assays detected cell proliferation. The cell cycle was detected by flow cytometry analysis. Wound healing and Transwell assays, respectively, estimated cell migration and invasion. The thiobarbituric acid reactive species (TBARS) method and BODIPY 581/591 C11 probe detected lipid peroxidation levels. FerroOrange staining estimated intracellular iron level. Western blot examined the expression of epithelial-mesenchymal transition (EMT) and ferroptosis-associated proteins. The human TFDB database predicted the binding of FOXA2 with the ACAT2 promoter, which was substantiated by ChIP and luciferase reporter assays. As a result, ACAT2 expression was increased in EC tissues and cells and associated with poor survival outcomes in EC patients. ACAT2 deletion might hinder EC cell proliferation, migration, invasion, and EMT while stimulating cell cycle arrest. Moreover, ACAT2 silencing promoted the ferroptosis of EC cells. Also, FOXA2 inactivated the transcription of ACAT2 through binding with the ACAT2 promoter. FOXA2 interference could promote the proliferation, migration, invasion, EMT, cell cycle, and inhibit the ferroptosis of ACAT2-silenced EC cells, which was partially reversed by the ferroptosis activator erastin. Conclusively, ACAT2 transcriptionally inactivated by FOXA2 might contribute to the malignant progression of EC via the inhibition of ferroptosis.
子宫内膜癌(EC)仍然是一种常见的妇科疾病,其发病率和死亡率持续上升。酰基辅酶A:胆固醇酰基转移酶2(ACAT2)在多种人类恶性肿瘤中通常被视为肿瘤促进因子。本研究旨在明确ACAT2在EC中的作用和机制,这方面尚未有相关报道。通过ENCORI和Kaplan-Meier plotter数据库分别分析了ACAT2在EC样本中的表达及预后意义。采用RT-qPCR和蛋白质免疫印迹法检测EC细胞中ACAT2和叉头框蛋白A2(FOXA2)的表达。使用CCK-8法、集落形成实验和EdU染色实验检测细胞增殖。通过流式细胞术分析检测细胞周期。分别采用伤口愈合实验和Transwell实验评估细胞迁移和侵袭能力。采用硫代巴比妥酸反应产物(TBARS)法和BODIPY 581/591 C11探针检测脂质过氧化水平。采用FerroOrange染色评估细胞内铁水平。蛋白质免疫印迹法检测上皮-间质转化(EMT)和铁死亡相关蛋白的表达。通过人类TFDB数据库预测FOXA2与ACAT2启动子的结合,并通过染色质免疫沉淀(ChIP)实验和荧光素酶报告基因实验进行验证。结果显示,ACAT2在EC组织和细胞中的表达增加,且与EC患者的不良生存结果相关。敲除ACAT2可能会阻碍EC细胞的增殖、迁移、侵袭和EMT,同时诱导细胞周期停滞。此外,沉默ACAT2可促进EC细胞的铁死亡。而且,FOXA2通过与ACAT2启动子结合使ACAT2转录失活。干扰FOXA2可促进ACAT2沉默的EC细胞的增殖、迁移、侵袭、EMT和细胞周期进程,并抑制其铁死亡,而铁死亡激活剂erastin可部分逆转这种作用。总之,被FOXA2转录失活的ACAT2可能通过抑制铁死亡促进EC的恶性进展。