龙葵糖蛋白的制备及其对MCF-7细胞毒性的作用.docVIP

  龙葵糖蛋白的制备及其对 MCF-7 细胞毒性 的作用# ， 5 10 15 20 25 30 35 40  摘要：目的 制备龙葵糖蛋白并观察其对 MCF-7 细胞的细胞毒作用。方法 从龙葵青果中分 离出龙葵粗多糖；Sevage 法除去游离蛋白；10%的 H2O2 脱色，95%乙醇沉淀，分离出龙葵 多糖。龙葵多糖经 DEAE-52 纤维素柱层析得多糖-蛋白复合物；采用 SDS电泳法检 测多糖-蛋白复合物是否为糖蛋白，并测定其相对分子量；MTT 法检测多糖-蛋白复合物对乳 腺癌细胞 MCF-7 的细胞毒作用的 IC50；再经 SephadexG-200 凝胶柱层析纯化；采用 MTT 法进行活性检测。结果 从龙葵青果中提取分离得到龙葵多糖粗提物，通过脱色、脱蛋白处 理得龙葵多糖。经 DEAE-52 纤维素柱层析得多糖-蛋白复合物，SDS电泳法检测，测 得其是相对分子质量为 30kD 和 25kD 的两种糖蛋白混合物。经 MTT 法测得其对乳腺癌细胞 MCF-7 的细胞毒活性，测得的 IC50 为 804.51μg/mL。然后经 SephadexG-200 凝胶柱层析纯 化得糖蛋白 A、B，并测的其 IC50 分别为 532.96μg/mL、613.91μg/mL。结论 龙葵多糖细胞 毒作用的物质基础是相对分子质量为 30kD 和 25kD 的两种糖蛋白。 关键词：龙葵多糖；糖蛋白；物质基础 中图分类号：R284 Study on the material basis of cytotoxic activity of polysaccharide of solanum nigrum Li Wenlan1,2, Ji Yubin1,2, Gao Shiyong1,2, Zou Xiang1,2, Ji Chenfeng1,2, Qu Zhongyuan1,2, Bai Jing1,2, Dong Fang1,2, Xu Lili1,2 (1. Center of Research and Development on Life Sciences and Environmental Sciences, Harbin University of Commerce, Harbin 150076; 2. Engineering Research Center of natural antineoplastic drugs，Ministry of Education, Harbin 150076) Abstract: Abstract: Objective To study the material basis of cytotoxic activity of polysaccharide of solanum nigrum. Methods Polysaccharide of solanum nigrum crude product was isolated from Solanum nigrum fruit, removed free protein by sevage method, decolored 10% H2O2 and prepared by 95% ethanol treatment. Polysaccharide-protein complex isolated by EDAE-52 was detected the relative molecular by SDS. MTT assay was used in detecting cytotoxicity of polysaccharide-protein complexes for MCF-7 cells in vitro. Then polysaccharide-protein complex was separated by SephadexG-200 and detected activity of MTT method. Results Polysaccharide-protein complexeswas isolated by DEAE-52 and was detected molecular wight by SDS. It suggested that polysaccharide-protein complexes that IC50 was 804.5g/mL was mixture of 30kD and 25kD. Glycoprotein A, B were gained by SephadexG-200, from Polysaccharide-protein complexes. IC50 of glycoprotein A w