艰难梭菌A毒素的抗肿瘤活性研究

目的　研究艰难梭菌A毒素的抗肿瘤活性。方法　从艰难梭菌VPI 1 0 4 63菌株脑心浸液培养物 ,经甲状腺球蛋白亲和层析和QSepharose层析提纯得到精制A毒素。通过应用苔盼蓝染色测定细胞脱落率 ,四氮噻唑蓝 (MTT)比色测定细胞存活率 ,[3H] Uridine(尿嘧啶 )测定细胞膜损伤 ,光学、荧光显微镜观察细胞的形态变化等方法测定比较A毒素对TPC 1细胞 (人甲状腺乳头癌细胞 )和Vero细胞 (非洲绿猴肾细胞 )的作用。结果　在A毒素的作用下 ,TPC 1细胞的增长抑制、凋亡指数、细胞脱落率及细胞膜损伤情况均高于Vero细胞 ,且这种作用具有时间和剂量依赖性。结论　A毒素对TPC 1细胞的杀伤作用远大于对Vero细胞的作用。以上结果对细菌毒素的开发利用、抗癌新药的探索研制具有潜在意义

The antitumor activity of clostridium difficile toxin A

AIM To examine the antitumor activity of clostridium difficile toxin A. METHODS Highly purified toxin A from clostridium difficile was obtained by bovine thyroglobulin affinity chromatography followed by ion exchange chromatography steps on Q sepharose. The antitumor activity of toxin A of clostridium difficile on TPC 1 cell line was studied with Vero cell line as the normal cell line. The estimating ways used in this study were trypan blue exclusion test, MTT calorimetric assay, membrane damage test using 3H Uridine and observation by optical, fluorescence microscopes. RESULTS Exposed to toxin A, the cell growth inhibition, apoptosis index, non adherent cells and membrane damage in TPC 1 cell line were much more great than that in vero cell line, and the effect was dependent upon the concentration and treating time. CONCLUSION The antitumor activity of toxin A on TPC 1 cells was much higher than that on vero cell line. The data are of potential importance for the development of toxin A and the exploration of antitumor drugs.