Abstract 4107: CircZFR-YTHDF3 complex drives lymph nodes metastasis in cervical cancer via facilitating FASN translation

Introduction: Lymph node metastasis is a key driver of poor outcomes in cervical cancer. Our study reveals that elevated fatty acid synthase (FASN) protein is closely linked to metastasis and reduced survival in cervical cancer, yet its regulatory mechanisms remained unclear. Methods: We compared the expression of FASN in the cervical cancer samples with or without lymph nodes, long or short disease-free survival (DFS) via qRT-PCR, western blot assay, and tissue array. We investigated and validated the combination between circZFR and YTH N6-methyladenosine RNA binding protein F3 (YTHDF3) protein via circRNA pulldown assay, mass spectrometry, and RNA immunoprecipitation (RIP) assay. The m6A sites on FASN mRNA were tested via MeRIP-qPCR. Protein co-immunoprecipitation (Co-IP) assay was used to validate the combination between YTHDF3 and eukaryotic translation initiation factor 4A3 (eIF4A3) protein. The siRNAs for YTHDF3 or FASN were transfected into HeLa, C33A, and Caski cells. Proliferation ability of cells was tested via CCK-8 and EdU assay. Migration and invasion ability of cells were tested via transwell assay. The function of circZFR on promoting lymph nodes metastasis was validated by animal assay. Results: The test in clinical samples showed that the overexpression of circZFR and FASN is associated with lymph nodes metastasis and shorter DFS of cervical cancer patients. The results of circRNA pulldown and RIP assay demonstrated the combination between circZFR and YTHDF3 protein. The results of MeRIP-qPCR assay validated the m6A sites in FASN mRNA. The results of Co-IP assay showed that upregulated circZFR could increase the combination between YTHDF3 and eIF4A3 proteins. The results of proliferation, migration, and invasion assay suggested that knocking down FASN will inhibit the proliferation, migration, and invasion of cervical cancer cells. The results of animal assay validated the promotion of lymph nodes metastasis by circZFR. Conclusions: We identified circular RNA circZFR as a crucial regulator, significantly enhancing FASN protein expression. CircZFR overexpression accelerates lymph node metastasis and shortens DFS in patients. Mechanistically, circZFR binds to the m6A reader protein YTHDF3, facilitating m6A recognition on FASN mRNA and recruiting the translation initiator eIF4A3, thereby boosting FASN translation. These findings establish circZFR as a pivotal driver of cervical cancer progression and highlight its inhibition as a promising therapeutic strategy. Citation Format: Mingyi Zhou, Yong Zhang, Lian He, Bo Gao, Francois Claret, George Calin, Danbo Wang. CircZFR-YTHDF3 complex drives lymph nodes metastasis in cervical cancer via facilitating FASN translation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 4107.