Safflower cell-derived EVs trigger multipronged inhibition in cancer cells

Medicinal plant-derived extracellular vesicles (EVs) have emerged as potential nano-regulators for inter-kingdom disease regulations. Safflower is a medicinal herb, and its EVs have been highlighted in previous studies for their potential application in cancer treatment. However, no research has been conducted to specifically assess their anti-cancer properties. Therefore, this study aims to investigate the anticancer therapeutic potential of EVs derived from Safflower cell suspension cultures (SFEVs). SFEVs were successfully isolated by ultrafiltration and characterized using transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), Fourier transform spectroscopy (FTIR), western blotting, and zeta potential measurement. This was followed by in vitro anticancer potential analysis on multiple cancer cell lines. Results revealed circular-to-oval vesicles averaging 90.2 nm in size, maintaining stable colloidal properties. Functional assessments showed significant, dose-dependent cytotoxicity in HepG2, HeLa, and A431 cancer cell lines. Moreover, transcriptomic analysis of A431 cells treated with SFEVs reveals extensive reprogramming in oncogenic signaling pathways, highlighting potential targets of SFEVs in cancers, indicating that SFEVs treatment initiates a multifaceted anticancer response with significant downregulation of the ID gene family (ID1, ID2, and ID3), which can potentially induce endoplasmic reticulum (ER) stress through DDIT3 (CHOP), and can possibly promote cell cycle arrest via CDKN1A. This molecular transformation is further characterized by increased expression of ERRFI1 and TNFRSF10B. In conclusion, this study clearly illustrates that SFEVs are a stable nano-composition with reliable characterization parameters and purity. It also establishes that SFEVs have potential for use in cancer research, and potential targets have been identified for future research, providing a path for further research.