葡萄球菌中毒性休克毒素的分离纯化及其体外抗肝癌活性分析

Purification and in vitro Anti-hepatocellular Carcinoma Activity of Toxic Shock Syndrome Toxin-1

采用固体膜进行TSST 1的产毒培养,然后从金葡菌产毒产物中经CM纤维素离子交换层析、SephadexG 75、SephacrylS 200三步纯化,成功地获得了纯度为95.3%的毒素,SDS PAGE呈现单一条带,与其对应抗血清有免疫学特异反应,不含核酸、糖和其它肠毒素等物质.与同类方法相比,该方法步骤少、快速、简便.对其体外抗肝癌活性进行了分析,首次证明即使TSST 1的浓度为10-8g L仍然对人肝癌细胞有显著抑制作用.

A simple and efficient three step procedure was developed to purify TSST1 after culturing the toxinproducing bacteria on membrane.The purification procedure involved ionexchange chromatography on CM cellulose,gel filtration on Sephadex G75 and Sephacryl S200.TSST1 with high purity was obtained.The purified toxin is a simple protein with a molecular weight of 22 kD as determined by SDSPAGE.Other material such as nuclear acid and staphylococcal enterotoxins are not contained in the purified products.Specific reaction of purified TSST1 is demonstrated by immunodiffusion test.Our improved purification procedure is simple, more rapid and convenient than previously reported methods.The antitumor activity of TSST1 was also analysed.It was demonstrated for the first time that even 10-8?g/L TSST1 can notably inhibit the tumor growth of human hepatocellular carcinoma in vitro.