Identification of ANC-3 as a Novel Therapeutic Candidate for Anaplastic Thyroid Cancer Through Drug Screening and Multi-Platform Validation.

Anaplastic thyroid carcinoma (ATC) is a rare but highly aggressive malignancy characterized by rapid progression, early metastasis, and extremely poor survival outcomes. Effective therapeutic options remain limited, highlighting the need for efficient and biologically relevant preclinical drug-discovery platforms. In this study, high-throughput compound screening using human ATC cell lines identified ANC-3 as a potential anticancer candidate. Its antitumor activity was evaluated through cytotoxicity and functional assays, zebrafish xenograft validation with live fluorescence imaging, colony-formation assays, and bulk RNA sequencing with pathway enrichment analyses. ANC-3 demonstrated consistent antitumor effects by significantly inhibiting cell viability, migration, invasion, and clonogenic survival, while also suppressing tumor growth in zebrafish xenograft models. Transcriptomic analyses revealed modulation of multiple oncogenic pathways, including MAPK, Ras, and NF-κB signaling. Collectively, these findings support zebrafish xenograft-based screening as a rapid and scalable platform for ATC drug discovery and suggest ANC-3 as a promising multi-pathway inhibitor warranting further preclinical development.