Hemorheology Center, Department of Medical Physics, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, China.
J Ethnopharmacol. 2010 Feb 17;127(3):702-8. doi: 10.1016/j.jep.2009.12.003. Epub 2009 Dec 5.
Gekko swinhonis Guenther has been used as an anti-cancer drug in traditional Chinese medicine for hundreds of years. Here we investigated the structural characterization and anti-cancer effects of sulfated polysaccharide-protein complex (GSPP) isolated from Gekko swinhonis Guenther.
The structure of GSPP was characterized by high performance liquid chromatography, gas chromatography, gas chromatography-mass spectrometry, beta-elimination reaction, and NMR spectroscopy. SMMC-7721 cells were used to assess the influence of GSPP on hepatocellular carcinoma. Cell proliferation and survival was determined by trypan blue exclusion assay. Cell migration was performed by wound-healing and transwell assay. The secretion of IL-8 was detected by an enzyme-linked immunosorbent assay kit. Flow cytometry was used to analyze intracellular calcium concentration, as well as cell cycle distribution and apoptosis. Confocal microscopy was used to assess the localization and configuration of actin filaments.
GSPP was chemically characterized as a sulfated polysaccharide-protein complex with O-glycopeptide linkages. Our results showed that GSPP inhibited the proliferation of SMMC-7721 cells and blocked cells in the S phase. No direct toxicity against cells was observed. Furthermore, GSPP inhibited the migration of SMMC-7721 cells with the reduction of intracellular calcium. Actin filaments were polymerized and accumulated in the cytoplasm of the treated cells, whereas the secretion of IL-8 was not significantly changed after GSPP exposure.
We describe an identified sulfated polysaccharide-protein complex, and demonstrate its direct effect on hepatocellular carcinoma cell migration via calcium-mediated regulation of the actin cytoskeleton reorganization.
蛤蚧已在传统中药中被使用数百年，作为一种抗癌药物。本研究旨在探索从蛤蚧中分离得到的硫酸多糖-蛋白复合物（GSPP）的结构特征及其抗癌作用。
采用高效液相色谱、气相色谱、气相色谱-质谱联用、β-消除反应和 NMR 光谱学等方法对 GSPP 的结构进行了表征。采用 SMMC-7721 细胞评估 GSPP 对肝癌的影响。通过台盼蓝排斥试验测定细胞增殖和存活情况。采用划痕愈合和 Transwell 试验检测细胞迁移。采用酶联免疫吸附试验试剂盒检测白细胞介素-8（IL-8）的分泌。采用流式细胞术分析细胞内钙离子浓度、细胞周期分布和细胞凋亡。采用共聚焦显微镜评估肌动蛋白丝的定位和构型。
GSPP 被化学表征为具有 O-糖肽键的硫酸多糖-蛋白复合物。结果表明，GSPP 抑制 SMMC-7721 细胞的增殖并将细胞阻滞在 S 期。未观察到对细胞的直接毒性。此外，GSPP 抑制 SMMC-7721 细胞的迁移，同时减少细胞内钙离子。处理后的细胞中肌动蛋白丝聚合并在细胞质中积累，而 GSPP 暴露后白细胞介素-8 的分泌没有明显变化。
本研究描述了一种已鉴定的硫酸多糖-蛋白复合物，并证明其通过钙介导的肌动蛋白细胞骨架重排调节直接作用于肝癌细胞迁移。
