蚯蚓组织中抑制肿瘤细胞生长物质的分离提取及其作用机制初探

A preliminary study on isolation of a tumor cell inhibiting active component from earthworm and its mechanism

目的 探索一条从蚯蚓中分离有效抑癌成分的途径 ,并对其作用机制进行初探。方法 选取赤子爱胜蚓 (EiseniaFoeti da) ,采用硫酸铵分段盐析、DEAE纤维素 DE5 2离子交换柱层析、SephadexG10 0凝胶过滤柱层析等方法分离纯化。MTT方法测定各分离步骤所得样品对于肿瘤细胞的抑制作用。流式细胞术、共聚焦激光扫描显微镜检测细胞凋亡。结果 蚯蚓组织抽提液经分离纯化后 ,得一蛋白组分。它可有效抑制BEL 74 0 2细胞 (人原发性肝癌细胞系 )、HeLa细胞 (人宫颈癌细胞系 )的生长。凝胶过滤测其分子量为 4 2 .2kD ,SDS PAGE显示两条带 (分别为 33和 10kD)。细胞凋亡检测结果证实此组分有促肿瘤细胞凋亡的作用。结论 对蚯蚓组织进行分离提取 ,可得到抑制实体瘤细胞生长的活性组分。它可引起肿瘤细胞的凋亡 ,凋亡机制有待进一步探索。

Objective To introduce a new method of isolation of an antitumor component from earthworm and to explore its mechanism. Methods The active component was isolated from earthworm (Eisenia Foetida) homogenate by a procedure involving (NH 4) 2SO 4 salting out, DEAE cellulose--DE52 anion exchange chromatography and Sephadex G100 gel filtration chromatography. The MTT test was used to assess the inhibitory effects of each samples obtained in isolation proceses on BEL-7402 (human hepatoma) cells and HeLa (cervix carcinoma) cells. Results An active component,a protein which could inhibit tumor cell growth was isolated .Its molecular weight was 42.2kD determined with gel filtration on Sephadex G100, where of the protein subunits were 33kD and 10kD estimated with sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Flow cytometry and focus laser scan microscopy proved that the active component could promote the apoptosis of tumor cells. Conclusion An active component that can inhibit tumor cells' growth was isolated from earthworm homogenate. It can promote the apoptosis of tumor cells and the mechanism is need to further study.