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.

Bladder cancer (BC) is one of the most common malignant tumors in the urinary system. Due to the poor prognosis and high mortality rate of the disease, it is urgent to develop new drugs with high efficacy and low toxicity to treat BC. Echinatin (Ecn) is a bioactive natural flavonoid oflicorice that has attracted special attention for its promising anti-tumor potential. Herein, we explored the inhibitory effects of Echinatin on BC cells and probed the possible molecular mechanism. We found that Ecnin vitro inhibited the proliferation, migration, and invasion, arrested the cell cycle at the G2/M phase, and promoted apoptosis in BC cells. Besides, Ecn had no notable cytotoxicity towards human normal cells. We subsequently confirmed that Ecn restrained xenograft tumor growth and metastasis of BC cells in vivo. Mechanistically, Ecn activated the p38 signaling pathway but inactivated the Wnt/β-catenin signaling pathway, while over-expression of β-catenin and the p38 inhibitor both attenuated the inhibitory effects of Ecn on BC cells. Remarkably, Ecn combined with cisplatin (DDP) or gemcitabine (Gem) had synergistic inhibitory effects on BC cells. In summary, our results validate that Ecn inhibits the tumor growth of human BC cells via p38 and Wnt/β-catenin signaling pathways. More meaningfully, our results suggest a potential strategy to enhance DDP- or Gem-induced inhibitory effects on BC cells by combining with Ecn.