Stylopine: A potential natural metabolite to activate vascular endothelial growth factor receptor 2 (VEGFR2) in osteosarcoma therapy

• Main Authors: Velayutham, Naveen Kumar, Thamaraikani, Tamilanban, Wahab, Shadma, Khalid, Mohammad, Gobinath, Ramachawolran, Abullais, Shahabe Saquib, Wong, Ling Shing, Mahendran, Sekar, Gan, Siew Hua, Ebenezer, Angel Jemima, Ravikumar, Mrinalini, Subramaniyan, Vetriselvan, Nur Najihah, Izzati Mat Rani, Wu, Yuan Seng *, Jeyabalan, Srikanth
• Format: Article
• Published: Frontiers Media 2023
• Subjects: QL Zoology; QP Physiology; RC Internal medicine
• Online Access: http://eprints.sunway.edu.my/2777/; https://doi.org/10.3389/fphar.2023.1150270
• Institution: Sunway University

Vascular endothelial growth factor (VEGF) signals cell survival, cell migration, osteogenesis, cell proliferation, angiogenesis, and vascular permeability by binding to VEGF receptor 2 (VEGFR-2). Osteosarcoma is the most common primary bone cancer, majorly affects young adults. Activation of VEGFR-2 signaling is a therapeutic target for osteosarcoma. The present study aimed to evaluate the potency of stylopine in regulation of the VEGFR-2 signaling pathway and its anti-tumour effect human MG-63 osteosarcoma cells. The in silico study on benzylisoquinoline alkaloids was carried out for analyzing and shortlisting of compounds using a virtual screening, Lipinski's rule, bioavailability graphical RADAR plot, pharmacokinetics, toxicity, and molecular docking studies. Among the benzylisoquinoline alkaloids, stylopine was selected and subjected to in-vitro studies against human MG-63 osteosarcoma cells. Various experiments such as MTT assay, EtBr/AO staining, mitochondrial membrane potential assessment, transwell migration assay, gene expression analysis by a quantitative real time polymerase chain reaction (qRT-PCR) method, SDS-PAGE followed by immunoblotting were performed to evaluate its anti-tumour effect as compared to standard axitinib. The MTT assay indicates that stylopine inhibits cell proliferation in MG-63 cells. Similarly, as confirmed by the EtBr/Ao staining method, the MMP assay indicates that stylopine induces mitochondrial membrane damage and apoptosis as compared to axitinib. Moreover, stylopine inhibits the VEGF-165 induced MG-63 cell migration by a trans-well migration assay. The immunoblotting and qRT-PCR analysis showed that stylopine inhibits the VEGF-165 induced VEGFR2 expression in MG-63 cells. It is concluded that stylopine has potential to regulate VEGFR2 and can inhibit osteosarcoma cells to offer a new drug candidate for the treatment of bone cancer in future.