A Di-Rhamnolipid (Rha2-C10-C10) Inhibits Colorectal Cancer Motility and Immune Evasion Through AKT-Associated Signaling and PD-L1 Regulation.

Colorectal cancer (CRC) progression is driven by metastatic potential, metabolic reprogramming, and immune evasion. In this study, we investigated the effects of a di-rhamnolipid, Rha2-C10-C10 (1), on CRC cell motility, energy metabolism, and immune-related signaling. Compound 1 exhibited minimal cytotoxicity in AGS, A549, and MDA-MB-231 cells, while modestly reducing viability in Caco2 cells at higher concentrations. Notably, it significantly suppressed invasion and migration, with the most pronounced effects observed in Caco2 cells. These effects were associated with downregulation of mesenchymal markers, including N-cadherin and transcription factors Snail and Slug, as well as matrix metalloproteinases (MMP2, MMP3, and MMP9), accompanied by increased TIMP2 expression. In addition, compound 1 attenuated metabolic activity by reducing the expression of key glycolytic regulators, including GLUT1, LDHA, and HK2. It also suppressed immune evasion-related factors such as PD-L1, IDO1, IDO2, and Galectin. Mechanistic investigations using BaP and the AKT inhibitor MK2206 suggest that the effects of compound 1 involve modulation of AKT-associated signaling while also engaging additional regulatory pathways. Collectively, these findings demonstrate that compound 1 suppresses colorectal cancer cell motility, metabolic activity, and immune evasion, highlighting its potential as a natural compound targeting multiple tumor-promoting processes.