摘要
目的探讨地鳖虫纤溶活性蛋白组分的提取及对肿瘤细胞的抑制作用。方法使用硫酸铵沉淀、DEAE离子交换层析和SephadexG75凝胶层析的方法，从地鳖虫体内提取出纤溶活性蛋白并检测其活性，研究其体外抗肿瘤的作用；并通过噻唑蓝（MTT）法和流式细胞术检测地鳖虫纤溶活性蛋白对2种肿瘤细胞（人食管癌细胞株Eca109和人宫颈癌细胞株HeLa）的增殖抑制及周期的影响。结果（1）地鳖虫体内存在具有纤溶活性的纤溶活性蛋白；（2）地鳖虫纤溶活性蛋白对肿瘤细胞的增殖抑制率与剂量和时间密切相关；（3）随着地鳖虫纤溶活性蛋白浓度的增加，G1期细胞数量明显减少，G2／M期的细胞数量有所增加。结论地鳖虫纤溶活性蛋白具有体外抑制肿瘤细胞的作用。
Objective To isolate and purify Eupolyphaga fibrinolyric protein （EFP） from Eupolyphaga sinesis Walker, and investigate the inhibited effects of EFP on tumour cells. Methods EFP was isolated and purified by ammonium precipitation, DEAE ion exchange chromatography and Sepha dexG-75 column chromatography from fresh Eupolyphaga sinesis Walker,and then its activity was de tected. The cells of human esophageal carcinoma strain （Ecal09） and human uterine cervix cancer （HeLa） strain were cultured and treated with EFP in order to detect the effect of EFP on cell generation and cell cycles with MTT and flow cytometric method. Results （1） There existed fibrinolytic protein which had activity in Eupolyphaga sinesis Walker; （2） EFP inhibited the growth of tumor cells in a dose- and time-dependent manner; （3） The number of cells in G1 phase decreased significantly, and in G2/M was elevated with the increase of the EFP concentration. Conclusion The results indicates that the EFP has antitumour effect in vitro.
出处
《国际检验医学杂志》
CAS
2007年第12期1088-1090,1093,共4页
International Journal of Laboratory Medicine
基金
广东省自然科学基金项目（04010984）
国家自然科学基金项目（30772739）
关键词
蜚蠊目
蛋白质类
分离和提纯
抗肿瘤药
Walkers
Proteins
Isolation ＆- purification
Antitumour agents
