掌叶半夏凝集素的分离纯化及其在体内外对小鼠肉瘤S180细胞的影响

目的:优化掌叶半夏凝集素的分离纯化方案,研究其在体内外对S180肿瘤细胞增殖的影响及细胞周期分布的特点。方法:经硫酸铵分级沉淀、热处理、DEAE离子交换层析和SDS-PAGE电泳等方法对掌叶半夏凝集素进行分离纯化,并用荧光光谱分析等生物化学方法研究其理化性质;MTT法检测其对S180细胞增殖的影响;流式细胞术检测其对S180细胞周期的影响,而动物实验观察其体内抑瘤效应。结果:掌叶半夏凝集素是一种非糖蛋白,分子质量为9 067.5 U,能特异性的凝集兔和鼠红细胞;与S180细胞作用24 h后有一定的抑制作用,而对其细胞周期(G2/M期)和凋亡率的影响不明显,体内抑瘤实验显示其平均抑瘤率仅为11.55%。结论:掌叶半夏凝集素体外能抑制S180细胞生长,却并非通过促进其细胞凋亡途径实现,而对S180荷瘤小鼠体内的肿瘤细胞杀伤作用弱且无明显的免疫激活作用。

Isolation and Purification of Pinellia Pedatisecta Lectin and Its Effects on Mouse Sarcoma S180 Cells in Vivo and in Vitro

Objective: To optimize the methods of isolation and purification of pinellia pedatisecta letin and to explore its effects on the proliferation and cell cycle distribution of mouse sarcoma S180 cells in vivo and in vitro.Methods: Pinellia pedatisecta letin can be purified gradually through ammonium sulphate fractionation,heat treatment,DEAE ionic exchange chromatographic analysis and SDS-PAGE electrophoresis.The physical and chemical character was analyzed by a series of methods.The effects of pinellia pedatisecta letin on the growth of S180 cells were studied by MTT assay by flow cytometry on the cell cycle phase distribution of S180 cells,and its antitumor effect in vivo was tested by animal experiment.Results: Pinellia pedatisecta letin could agglutinate the fresh erythocytes of rabbit and mouse,of which the molecular weight is 9 067.5 U,but surprisingly it contains no sugar chain.After 24 hours,the growth of S180 cells was inhibited,but it displayed no effect on the cell cycle phase distribution.Animal experiment suggested that the average antitumor rate was only 11.55%.Conclusion: Pinellia pedatisecta letin could inhibit the proliferation of S180 cells,but the effect of which on inducing cell apoptosis and regulating the cell cycle is not obvious.It hadn't antitumor activity in vivo and could not improve the immunity activity effectively.