摘要
目的从长白山岩栖蝮蛇蛇毒中分离纯化1种小分子多肽Saxis,测定理化性质,研究其对肿瘤细胞的生长抑制作用。方法利用有机溶剂沉淀和Sephadex G-50S、ephadex G-25凝胶过滤色谱等方法分离纯化长白山栖岩蝮蛇蛇毒中的1种小分子多肽Saxis;采用Tricine-SDS-PAGE鉴定;利用Bradford蛋白质测定法测定蛋白质浓度;运用MTT比色法检测该小分子多肽对宫颈癌细胞Hela、肝癌细胞SMMC-7721和胃腺癌细胞SGC-7901的生长抑制作用。结果该小分子多肽Saxis的相对分子质量约为5 200,它能抑制Hela、SMMC-7721和SGC-7901细胞的增殖,并且对这3种肿瘤细胞的抗增殖作用具有药物剂量依赖关系,24 h的IC50分别为66.3,54.5,62.2μg/mL。结论利用有机溶剂沉淀和凝胶过滤色谱法可以从长白山岩栖蝮蛇蛇毒中分离纯化1种小分子多肽Saxis,其对多种肿瘤细胞的生长有抑制作用。
Purpose To purify a small peptide from snake venom of Gloydius saxatilis, to identify its physical and chemical properties and to investigate the inhibitory effect of the small peptide on the proliferation of tumor cells. Methods The small peptide was purified by gel filtration chromatography, the molecular weight was estimated by Tricine-SDS-PAGE and the concentration was determined by Bradford Method. Its effect on the proliferation of SMMC-7721, Hela and SGC-7901 cells was analyzed by MTT assay. Results A small peptide was purified from snake venom of Gloydius saxatilis by gel filtration chromatography, and it was identified as a single component by Tricine-SDS-PAGE. Its moleular weight was about 5.2 kDa. The small peptide can dose-dependently inhibit the proliferation of SMMC-7721, Hela and SGC-7901 cells and the ICs0 for 24 h culure were 66.3,54.5 and 62.2 μg/mL respectively. Conclusion A small peptide can be obtained by means of organic solvent precipitation and gel filtration, which exhibits strong effect on inhibiting the proliferation of cultural tumor cells line in vitro.
出处
《中国生化药物杂志》
CAS
CSCD
北大核心
2009年第4期217-220,225,共5页
Chinese Journal of Biochemical Pharmaceutics
基金
吉林省科技厅项目(20060564)
关键词
蛇毒
多肽
分离纯化
肿瘤细胞
snake venom
peptide
purification
tumor cells
