Gao Fei, Wang Xiangjun, Li Zhao, Zhou Aicun, Tiffany-Castiglioni Evelyn, Xie Lijun, Qian Yongchang
Department of Traditional Chinese Medicine, Zhejiang A&F University, Lin'An, Zhejiang 311300, P.R. China.
Nurturing Station for the State Key Laboratory of Subtropical Silviculture, School of Forestry and Biotechnology, Zhejiang A&F University, Lin'An, Zhejiang 311300, P.R. China.
Oncol Lett. 2017 Jul;14(1):15-22. doi: 10.3892/ol.2017.6160. Epub 2017 May 12.
Secretion of granular glands from the skin of amphibians is a fascinating resource of active substances, particularly for cancer therapy in clinical practice of Traditional Chinese Medicine. A variety of anti-tumor peptides have been isolated from different toads and frogs; however, no anti-tumor peptides are reported in toad venom of . Firstly, soluble fraction from fresh toad venom (FTV) was compared with that from dried toad venom (DTV), using HPLC analysis. It was revealed that FTV has a different HPLC chromatography compared with DTV. Soluble fraction of FTV is more toxic to SH-SY5Y cells than that of DTV, as evaluated by MTT assay. Secondly, it was identified that protein components from soluble fractions of FTV and DTV possess different patterns by SDS-PAGE analysis, and proteins from FTV are also more toxic than that from DTV. Thirdly, an immobilized basic fibroblast growth factor (bFGF) affinity column was used to isolate bFGF-binding components from soluble fraction of FTV, and it was identified that bFGF-binding components prohibited bFGF-dependent neurite growth of SH-SY5Y cells. Finally, it was identified that bFGF-binding components activated apoptosis via upregulation of caspase-3 and caspase-8 expression in SH-SY5Y cells. These data suggest that FTV contains active components that interact with bFGF and activate apoptosis in SH-SY5Y cells.
两栖动物皮肤颗粒腺的分泌物是一种引人入胜的活性物质来源，尤其在中医临床实践中对癌症治疗具有重要意义。人们已从不同蟾蜍和青蛙中分离出多种抗肿瘤肽；然而，尚未有关于[蟾蜍名称缺失]蟾蜍毒液中抗肿瘤肽的报道。首先，通过高效液相色谱分析比较了新鲜蟾蜍毒液（FTV）和干蟾蜍毒液（DTV）的可溶部分。结果显示，FTV与DTV具有不同的高效液相色谱图谱。通过MTT法评估发现，FTV的可溶部分对SH-SY5Y细胞的毒性比DTV的更强。其次，通过SDS-PAGE分析确定，FTV和DTV可溶部分的蛋白质成分具有不同模式，且FTV中的蛋白质毒性也比DTV中的更强。第三，使用固定化碱性成纤维细胞生长因子（bFGF）亲和柱从FTV的可溶部分中分离bFGF结合成分，并确定bFGF结合成分可抑制SH-SY5Y细胞中bFGF依赖的神经突生长。最后，确定bFGF结合成分通过上调SH-SY5Y细胞中caspase-3和caspase-8的表达来激活细胞凋亡。这些数据表明，FTV含有与bFGF相互作用并在SH-SY5Y细胞中激活细胞凋亡的活性成分。
