蚯蚓中抗肿瘤蛋白组分的提取分离及其抗肿瘤活性

以赤子爱胜蚓为原料 ,通过蛋白质的丙酮沉淀和凝胶过滤 ,分离得到一组抗肿瘤活性蛋白成分 (简称蚯蚓抽提物 ) ;这组成分富含Fe、Zn、Cu以及Se等微量元素 ,蛋白含量为 6 0 4 3± 2 36 %(n =5 ) .体外实验中 ,通过MTT法和SRB法测定了蚯蚓抽提物对多种人癌细胞株 (HCT 116、SY5Y、K5 6 2、MGc80 3和HeLa)以及正常细胞株 (HEK2 93和COS 7)的抑制杀伤率 .结果表明 ,蚯蚓抽提物对癌细胞的杀伤有一定的选择性 ,受试人癌细胞达到 5 0 %生长抑制率所需作用浓度约 6 0～110mg L ;但是 ,10 0℃煮沸 5min后 ,该抑制活性完全消失 .通过纤维蛋白平板法 ,测得蚯蚓抽提物同时具有纤溶酶和纤溶酶原激活酶的活性 .体外测得丝氨酸蛋白酶抑制剂aprotinin和PMSF能显著抑制蚯蚓抽提物的细胞杀伤活性 .体内实验中 ,蚯蚓抽提物能有效延长荷瘤 (S180 )小鼠的生存时间 ,并使其身体机能得到明显改善 .腹膜内注射剂量为 2 8mg kg和 36mg kg时 ,生命延长率分别为135 3%和 12 3 5 % ,和标准药物 (环磷酰胺 )治疗后结果 (76 5 % )相比 ,有显著性差异

Extraction and Isolation of the Anti-tumor Protein Components from Earthworm (Eisenia fetida andrei) and the Anti-tumor Activity

A group of anti tumor protein components ( Eisenia fetida extract, EE) was extracted and isolated from earthworm Eisenia fetida andrei by acetone sedimentation and gel filtration. The protein content of EE was 60.43±2 36% ( n =5). EE was also rich in trace elements, such as Zn, Cu, Fe and Se. In vitro the concentrations of EE required for 50% growth inhibition of different human tumor cell strains (HCT 116, SY5Y, K562, MGc803 and HeLa) were between 60 and 110 mg/L. And the inhibition of normal cell strains, HEK293 and COS 7, by EE were much weaker than those of most human tumor cell strains tested. However, the cell killing activity of EE was completely diminished after boiling for 5 min. By fibrin plate assay EE was found to be fibrinolytic as well as plasminogen activating. The cell killing activity of EE in vitro could be inhibited by serine protease inhibitors, aprotinin and phenylmethyl sulfonyl fluoride (PMSF). EE (28 mg/kg and 36 mg/kg, i.p.) in vivo were found to prolong the life span of ascites tumor (S180) bearing mice by 135 3% and 123 5%, respectively, and even to improve their physical conditions. Meanwhile, EE was found to be low toxic as compared with the cyclophosphamide treatment (76 5%).