brucine and Carcinoma--Ehrlich-Tumor

In this study, we investigated the mechanism of brucine in tumor angiogenesis. We found that brucine inhibits VEGF-induced cell proliferation, chemotactic motility, and the formation of capillary-like structures in HUVECs in a dose-dependent manner. Brucine suppresses VEGF- induced p-VEGFR2 kinase activity and inhibits neovascularization in vivo. Brucine inhibits the downstream protein kinases of VEGFR2, including Src, FAK, ERK, AKT and mTOR. And further downregulates levels of VEGF, NO, IL-6, IL-8, TNF-α and IFN-γ in HUVECs. Taken together, our study suggests that brucine potently suppresses angiogenesis by targeting VEGFR2 activation and may be a viable drug candidate in anti-angiogenesis and anti-cancer therapies.

Topics: Angiogenesis Inhibitors; Animals; Carcinoma, Ehrlich Tumor; Cell Proliferation; Cell Survival; Cells, Cultured; Chemotaxis; Cytokines; Dose-Response Relationship, Drug; Female; Gene Expression Regulation; Human Umbilical Vein Endothelial Cells; Humans; Male; Mice; Neovascularization, Physiologic; Nitric Oxide; Phosphorylation; Plants, Medicinal; Protein Kinase Inhibitors; Rats; Rats, Sprague-Dawley; RNA, Messenger; Signal Transduction; Strychnine; Strychnos nux-vomica; Time Factors; Tissue Culture Techniques; Tumor Burden; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor Receptor-2

Brucine (BRU), a natural plant alkaloid is reported to possess cytotoxic and antiproliferative activities. In this study we aimed to investigate its in vitro and in vivo antitumor and antiangiogenic effects.. Cell proliferation and viability was assessed using microculture tetrazolium tests (MTT). As predictive markers we determined intracellular levels of vascular endothelial growth factor (VEGF), Interleukin-12 (IL-12), tumor necrosis factor (TNF-α), caspase-3, -8 and -9 by ELISA and enzymatic activity assays. In addition, anti-VEGF neutralization effect was evaluated to assess whether it could result in augmented anticancer efficacy than the single agent. Antitumor activity was evaluated against Ehrlich ascites and solid tumor models. 15×10(6) EAC cells were implanted intraperitoneally (i.p., ascites tumor) and subcutaneous (s.c., solid tumor) in Swiss albino mice. Mice with established tumors received brucine i.p. at 12.5, 25, and 50mg/kg for 14days in ascites tumor and 50mg/kg in solid tumor for 30days. Tumor volume, cell viability, angiogenic, anti-angiogenic, anti-inflammatory factors and antioxidant parameters were determined. Immunohistochemistry analysis for VEGF and CD-31 was also performed.. BRU produced time and dose-dependent inhibition of MCF-7 in vitro and EAC tumors in vivo. The anti-angiogenic effects were accompanied with decreased VEGF and TNF-α and increased IL-12 expression. BRU reduced peritoneal angiogenesis and microvessel density in vivo.. Our findings suggest that BRU possesses antitumor and anti-angiogenic activities in vitro and in vivo. The above results showed that BRU can be used as a potential anticancer agent as an antimetastatic and anti-angiogenic agent.

Topics: Animals; Antineoplastic Agents; Antioxidants; bcl-2-Associated X Protein; Blood Cell Count; Carcinoma, Ehrlich Tumor; Caspases; Cell Line, Tumor; Dose-Response Relationship, Drug; Enzyme Activation; Female; Humans; Immunohistochemistry; Interleukin-12; Mice; Neoplasm Transplantation; Neovascularization, Pathologic; Oxidative Stress; Strychnine; Survival Analysis; Tetrazolium Salts; Thiazoles; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A