Zhao Lingqin, Song Qian, Zheng Chao, Sun Wei, Chen Yaqing
Department of Gynecologic Oncology, Zhejiang Cancer Hospital, Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences, Hangzhou, Zhejiang, China.
Department of Gynecologic Oncology, Taizhou Cancer Hospital, Wenling, Zhejiang, China.
J Biochem Mol Toxicol. 2025 Feb;39(2):e70140. doi: 10.1002/jbt.70140.
TWIST1 is aberrantly expressed in ovarian cancer (OC). MFAP2 is a downstream target of TWIST1, and we previously found MFAP2 facilitated OC development by activating FOXM1/β-catenin. We planned to investigate the mechanisms of TWIST1 in OC. GEPIA (a database for gene expression analysis) and UALCAN (a database containing comprehensive cancer transcriptome and clinical patient data) investigated TWIST1's connection to MFAP2 and patient survival in ovarian serous cystadenocarcinoma (OV). Human OC cells (A2780 and CAOV3) were transfected with si-TWIST1, oe-TWIST1, oe-MFAP2, or si-TWIST1 + oe-MFAP2. Cellular apoptosis, viability, migration, and invasion were detected. TWIST1, MFAP2, FOXM1, and β-catenin protein expressions were tested. Dual-luciferase and ChIP-qPCR validated the correlation between MFAP2 and TWIST1. Moreover, OC mice were established by injecting OC cells subcutaneously. The pathology, apoptosis, as well as Ki67, TWIST1, MFAP2, FOXM1, and β-catenin protein levels of tumors were assessed. TWIST1 expression positively correlated with MFAP2 expression, but negatively related to patients' survival in OV. TWIST1 overexpression promoted malignant behaviors, and increased MFAP2, FOXM1, and β-catenin protein levels for OC cells. TWIST1 knockdown exhibited the opposite trend. In vivo, TWIST1 knockdown disrupted tissue structure, induced apoptosis, decreased Ki67, TWIST1, MFAP2, FOXM1, and β-catenin protein levels in tumor. Interestingly, MFAP2 overexpression reversed the effects of TWIST1 knockdown in vitro and in vivo. Additionally, dual-luciferase and ChIP-qPCR confirmed MFAP2 was a downstream target for TWIST1 in OC. TWIST1 regulated FOXM1/β-catenin to promote the growth, migration, and invasion of OC cells by activating MFAP2, indicating that targeting TWIST1 may be effective for treating OC.
TWIST1在卵巢癌（OC）中异常表达。MFAP2是TWIST1的下游靶点，我们之前发现MFAP2通过激活FOXM1/β-连环蛋白促进OC发展。我们计划研究TWIST1在OC中的作用机制。GEPIA（一个基因表达分析数据库）和UALCAN（一个包含全面癌症转录组和临床患者数据的数据库）研究了TWIST1与MFAP2的关联以及卵巢浆液性囊腺癌（OV）患者的生存率。用人OC细胞（A2780和CAOV3）转染si-TWIST1、oe-TWIST1、oe-MFAP2或si-TWIST1 + oe-MFAP2。检测细胞凋亡、活力、迁移和侵袭情况。检测TWIST1、MFAP2、FOXM1和β-连环蛋白的蛋白表达。双荧光素酶和染色质免疫沉淀定量PCR（ChIP-qPCR）验证了MFAP2与TWIST1之间的相关性。此外，通过皮下注射OC细胞建立OC小鼠模型。评估肿瘤的病理学、凋亡情况以及Ki67、TWIST1、MFAP2、FOXM1和β-连环蛋白的蛋白水平。TWIST1表达与MFAP2表达呈正相关，但与OV患者的生存率呈负相关。TWIST1过表达促进恶性行为，并增加OC细胞中MFAP2、FOXM1和β-连环蛋白的蛋白水平。TWIST1敲低呈现相反趋势。在体内，TWIST1敲低破坏组织结构，诱导凋亡，降低肿瘤中Ki67、TWIST1、MFAP2、FOXM1和β-连环蛋白的蛋白水平。有趣的是，MFAP2过表达在体外和体内均逆转了TWIST1敲低的作用。此外，双荧光素酶和ChIP-qPCR证实MFAP2是OC中TWIST1的下游靶点。TWIST1通过激活MFAP2调节FOXM1/β-连环蛋白，以促进OC细胞的生长、迁移和侵袭，这表明靶向TWIST1可能对治疗OC有效。
