Effects of Gekko sulfated polysaccharide-protein complex on human hepatoma SMMC-7721 cells: Inhibition of proliferation and migration
Effects of Gekko sulfated polysaccharide-protein complex on human hepatoma SMMC-7721 cells: Inhibition of proliferation and migration
复制标题
Gekko硫酸化多糖蛋白复合物对人肝癌SMMC-7721细胞的增殖和迁移抑制作用
DOI：
10.1016/j.jep.2009.12.003
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发表时间：
2010-02-17
影响因子：
5.4
通讯作者：
Wen, Zong-Yao
中科院分区：
文献类型：
--
作者：
Chen, Dan;Yao, Wei-Juan;Wen, Zong-Yao
研究目的：在传统中医中，猪尾草作为抗癌药物已有数百年的历史。本文研究了虎斑鳖硫酸酸化多糖蛋白复合物（GSPP）的结构特征及其抗癌作用。材料与方法：采用高效液相色谱法、气相色谱法、气相色谱-质谱法、β -消除反应法和核磁共振波谱法对GSPP的结构进行表征。采用SMMC-7721细胞评价GSPP对肝细胞癌的影响。台盼蓝排斥法测定细胞增殖和存活。通过伤口愈合和transwell实验进行细胞迁移。采用酶联免疫吸附测定试剂盒检测IL-8的分泌。流式细胞术检测细胞内钙浓度、细胞周期分布及凋亡情况。使用共聚焦显微镜评估肌动蛋白丝的定位和结构。结果：GSPP的化学性质为巯基多糖-蛋白复合物，具有o -糖肽键。结果表明，GSPP对SMMC-7721细胞的增殖有抑制作用，使细胞处于S期。未观察到对细胞的直接毒性作用。此外，GSPP通过降低细胞内钙含量抑制SMMC-7721细胞的迁移。GSPP处理后，肌动蛋白丝在细胞质中聚合和积累，而IL-8的分泌没有明显变化。结论：我们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组调节对肝癌细胞迁移的直接影响。2009爱思唯尔爱尔兰有限公司版权所有。
Aim of the study: 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.Materials and methods: 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 immumosorbent 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.Results: 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.Conclusion: 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. (C) 2009 Elsevier Ireland Ltd. All rights reserved.