School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi Province, China.
J Cell Mol Med. 2013 Oct;17(10):1316-24. doi: 10.1111/jcmm.12108. Epub 2013 Aug 2.
Angiogenesis plays an important role in colon cancer development. This study aimed to demonstrate the effect of brucine on tumour angiogenesis and its mechanism of action. The anti-angiogenic effect was evaluated on the chicken chorioallantoic membrane (CAM) model and tube formation. The mechanism was demonstrated through detecting mRNA and protein expressions of VEGFR2 (KDR), PKCα, PLCγ and Raf1 by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot (WB), as well as expressions of VEGF and PKCβ and mTOR by ELISA and WB. The results showed that brucine significantly reduced angiogenesis of CAM and tube formation, inhibited the VEGF secretion and mTOR expression in LoVo cell and down-regulated the mRNA and phosphorylation protein expressions of KDR, PKCα, PLCγ and Raf1. In addition, the effects of brucine on KDR kinase activity, viability of LoVo cell and gene knockdown cell were detected with the Lance™ assay, WST-1 assay and instantaneous siRNA. Compared to that of normal LoVo cells, the inhibition on proliferation of knockdown cells by brucine decreased significantly. These results suggest that brucine could inhibit angiogenesis and be a useful therapeutic candidate for colon cancer intervention.
血管生成在结肠癌的发展中起着重要作用。本研究旨在展示白屈菜红碱对肿瘤血管生成的影响及其作用机制。采用鸡胚尿囊膜（CAM）模型和管形成实验评估抗血管生成作用。通过逆转录-聚合酶链反应（RT-PCR）和 Western blot（WB）检测 VEGFR2（KDR）、PKCα、PLCγ 和 Raf1 的 mRNA 和蛋白表达，以及 ELISA 和 WB 检测 VEGF 和 PKCβ和 mTOR 的表达，来证明机制。结果表明，白屈菜红碱可显著抑制 CAM 血管生成和管形成，抑制 LoVo 细胞 VEGF 的分泌和 mTOR 的表达，并下调 KDR、PKCα、PLCγ 和 Raf1 的 mRNA 和磷酸化蛋白表达。此外，还通过 Lance™ assay、WST-1 assay 和瞬时 siRNA 检测了白屈菜红碱对 KDR 激酶活性、LoVo 细胞活力和基因敲低细胞的影响。与正常 LoVo 细胞相比，白屈菜红碱对基因敲低细胞的增殖抑制作用明显降低。这些结果表明，白屈菜红碱可抑制血管生成，是结肠癌干预的一种有价值的治疗候选药物。
