Chondrosarcoma is recognized as the second most common form of primary bone cancer originating from cartilage. Antrodia cinnamomea is the source of the triterpenoid Antcin K, renowned for its immunomodulatory and anti-inflammatory properties. However, the therapeutic function of Antcin K on chondrosarcoma remains unclear. Our investigation found that Antcin K reduces migration and invasion in two chondrosarcoma cell lines. Finally, our study provided evidence of Antcin K as a novel candidate for inhibiting chondrosarcoma motility by inhibiting MMP-7 expression through suppressing the PI3K, Akt, mTOR, and NF-κB signaling cascades. Overall design: To discover differentially expressed mRNAs modulated by Antcin K, JJ012 cells were treated with Antcin K (10 μM) for 24 h in the culture medium, followed by mRNA sequencing.