糙皮侧耳糖蛋白的性质及体外抗肿瘤活性研究

Studies on the properties and antitumor activity in vitro of the glycoprotein from pleurotus ostreatus

目的 :对糙皮侧耳真菌提取液进行纯化 ,并对纯化组分的理化性质及体外抗肿瘤作用进行研究。方法 :用离子交换、凝胶过滤进行纯化 ,采用SDS PAGE测定蛋白分子量 ,用氨基酸自动分析仪测定氨基酸的组成 ,通过福林 酚法测定其蛋白和多糖的含量 ,用薄层层析确定多糖的组成 ,将POGP直接与S180 腹水瘤细胞及小鼠正常细胞液混合 ,观察其活性。结果 :纯化得到对肿瘤细胞具有细胞毒作用的糖蛋白 (POGP) ,SDS PAGE显示POGP为单一色带 ,其分子量为 1 0 2 .3KD ,氨基酸组成中鸟氨酸、精氨酸含量较高 ,蛋白与多糖的含量分别为 78.34 %和 1 6.53% ,其多糖主要由甘露糖组成 ,POGP对小鼠正常细胞均无作用 ,对S180 腹水瘤细胞具有细胞毒作用 ,甘露糖可抑制POGP的抗瘤活性。结论 :POGP具有特异性抗肿瘤作用 。

OBJECTIVE:To purify the extract from pleurotus ostreatus,and study on the physicochemical properties and antitumor activity in vitro of the purified component.METHOD:The ion exchange and gel filteration chromatograph were used.The molecular weight was determined with SDS PAGE.The composition of animo acids were determined with animo acids automatic analysis instrument.The folin phenol method and the sulfuric acid phenyphenol method were used to determine the contents of protein and polysaccharide.The compositions of polysaccharide were determined with thin layer chromatograph.The purified component was added to the S 180 ascite tumor cells and the mouse′s normal cells solution to test the activity.RESULTS:The glycoprotein POGP with cytotoxicity to tumor cells was separated from plrurotus ostreatus.SDS PAGE showed that POGP had a single band,and its MW was 102.3KD.The contents of orns and args in POGP were high.There were 78.34% of protein and 16.53% of polysaccharide in POGP.The polysaccharide was mainly composed of mannose.POGP had cytotoxocity to S 180 ascite tumor cells and no activity to mouse′s normal cells.The mannose could inhibit the cytotoxicity of POGP.CONCLUSION:POGP has specific antitumor activity and need further researching.