Background: Quercetin, a naturally occurring flavonoid with recognized antitumor properties, has limited therapeutic applicability due to its low water solubility and bioavailability. To address these issues, a new quercetin derivative, ANL3, was created with superior pharmacokinetic and physicochemical characteristics. Methods and Results: In vitro investigations showed that ANL3 effectively inhibited the proliferation, invasion, and migration of human osteosarcoma cells (MG-63 and SaOS-2) with lower IC 50 values than quercetin. Transcriptomic analysis and molecular testing revealed the FOXO1A–NDRG2– superoxide dismutase 2 ( SOD2 ) axis as a critical mechanistic route. ANL3 directly interacted with FOXO1A , increasing its phosphorylation without decreasing total protein expression, upregulating NDRG2 , and downregulating SOD2 , resulting in increased reactive oxygen species (ROS) buildup and endoplasmic reticulum (ER) stress. This cascade reduced the epithelial–mesenchymal transition and slowed osteosarcoma growth. In vivo , ANL3 therapy decreased tumor volume, increased survival in naked mice, and had little systemic toxicity. Ultrasound Augmentation: Based on these findings, low-intensity focused ultrasound was used to increase ANL3 delivery and intracellular activation. Ultrasound exposure increased cellular uptake, boosted local ROS production, and amplified ER stress signaling via the FOXO1A-NDRG2-SOD2 pathway. Conclusions: These findings show that ultrasound-augmented ANL3 treatment is a viable biotherapeutic method for osteosarcoma because it improves ER stress-mediated tumor suppression while reducing systemic adverse effects. This article proposes a mechanistic framework for incorporating ultrasound-mediated medication activation into precision cancer biotherapy.
Qian et al. (Thu,) studied this question.