Osteosarcoma (OS) is a highly aggressive bone malignancy with limited treatment options and frequent chemoresistance. Yanghe Decoction (YHD), a traditional Chinese medicine formula, has demonstrated anti-tumor potential, but its mechanisms in OS remain unclear. In this study, we employed a network pharmacology approach to identify 67 active components and 101 OS-related targets of YHD, with core targets including AKT1, TP53, MAPK14, and CASP3, mainly enriched in the PI3K/AKT and MAPK signaling pathways. Molecular docking confirmed strong binding affinities between representative compounds and these targets. Functional experiments revealed that YHD inhibited OS cell proliferation, migration, and invasion, and promoted apoptosis by elevating intracellular reactive oxygen species levels and inducing mitochondrial dysfunction. Mechanistically, YHD suppressed the PI3K/AKT pathway while activating p38 MAPK signaling. Importantly, YHD enhanced the sensitivity of OS cells to cisplatin, demonstrating a synergistic inhibitory effect in vitro and in an orthotopic OS mouse model. These findings suggest that YHD exerts its anti-osteosarcoma effects via reactive oxygen species-mediated mitochondrial disruption and pathway modulation, and may serve as a promising adjuvant to conventional chemotherapy.

Osteosarcoma (OS) is a highly aggressive tumor with a propensity for early metastasis. Current treatment methods, such as chemotherapy, often bring significant side effects, affecting patients’ quality of life. Veratramine (VER), an alkaloid derived from the American lily plant, has shown potential in cancer treatment. This study looks at the effects and mechanisms of VER on osteosarcoma. VER’s impact was assessed using a variety of procedures, including crystal violet staining, the CCK-8 assay, and the colony formation assay, which measured cell proliferation. Wound healing assay and transwell assay were employed to evaluate the migration and invasion of osteosarcoma cells. Hoechst33258 staining, flow cytometry, and transmission electron microscopy were used to investigate apoptosis. Protein expression was assessed using western blotting and immunofluorescence. Blood tests and hematoxylin-eosin staining were used to establish VER’s in vivo safety, and its effectiveness was proven using an orthotopic tumor model. The results showed that VER greatly decreased osteosarcoma cell growth, migration, and invasion while inducing apoptosis. Animal tests confirmed these findings, confirming VER’s high efficacy and safety in vivo. VER might function by inhibiting the PI3K/AKT signaling pathway. To sum up, VER shows promise in treating osteosarcoma by exhibiting significant anti-tumor activity in laboratory and animal studies, likely through the regulation of the PI3K/AKT signaling pathway.

Bone morphogenetic protein 9 (BMP9) has remarkable potential to induce the differentiation of mesenchymal stem cells (MSCs) towards the osteoblastic lineage. Additionally, research suggests that certain growth factors have the ability to potentiate BMP9-induced osteogenic differentiation of MSCs. Sonic Hedgehog (Shh) plays an indispensable role in the regulation of skeletal development. The objective of this research was to assess the potential influence of Shh on BMP9-induced osteogenic differentiation of MSCs. Our findings indicated that Shh effectively enhanced BMP9-induced early and late osteogenic differentiation of MSCs, and increased BMP9-induced expression/transcriptional activity of osteogenesis-related transcription factors. Besides, it was observed that Shh promoted BMP9-induced ectopic bone formation of MSCs in vivo. Moreover, BMP9 was able to facilitate the repair of bone defects in rats, while Shh further accelerated this reparative process. Mechanistically, Shh enhanced the activation of the Smad1/5/8 signaling pathway which was induced by BMP9. Furthermore, GANT-61, an inhibitor of Gli1 and Gli2, attenuated the enhancing effect of Shh on BMP9-induced osteogenic differentiation of MSCs. Collectively, the co-administration of BMP9 and Shh may present a promising therapeutic approach for the treatment of fracture nonunion, delayed fracture healing, and bone defects.