重组人内皮抑素提取方法的优化及其对膀胱癌的抑制作用  

Optimization of extracting method and inhibitory action of recombinant human endostatin in bladder cancer

作　　者：任明华[1] 倪少滨[1] 陈起引[1] 徐万海[2] 王长林[1] 付宜鸣[1] 焦治兴[1] 麻立[1] 赵忠山[1] 刘兴汉[3] 

机构地区：[1]哈尔滨医科大学第一临床医学院泌尿外科,黑龙江哈尔滨150001 [2]哈尔滨医科大学第四临床医学院泌尿外科 [3]哈尔滨医科大学生物化学与分子生物学教研室

出　　处：《中华医学杂志》2009年第46期3285-3288,共4页National Medical Journal of China

基　　金：黑龙江省青年科学技术专项基金（QC05C81）;黑龙江省卫生厅基金（2005-159）;教育厅基金（11521188）.

摘　　要：目的摸索重组人内皮抑素（rhES）蛋白的最佳表达条件。探索内皮抑素（ES）对膀胱癌实体肿瘤及细胞系的抑制作用。方法分别从不同异丙基-β-D硫代半乳糖苷（IPTG）浓度、加IPTG时机、表达时间等几个方面对rhES的蛋白表达条件进行摸索和优化。活性检测：（1）动物实验：观察内皮抑素对裸鼠皮下移植瘤的生长情况的影响。（2）细胞实验：MTT法、肿瘤细胞生长曲线观察内皮抑素对T-24膀胱癌细胞系的作用。结果rhES在大肠杆菌中的表达量占总菌体蛋白的58.65％，纯化后的蛋白纯度为96.22％。动物实验：rhES的抑瘤率为66.8％。细胞实验：MTT实验中rhES的半数抑制浓度IC50=22μg／ml；肿瘤细胞生长曲线表明第7天时rhES组的细胞数比对照组减少了58.75％。结论本实验所优化的最佳表达条件可获得高表达、高活性的rhEs蛋白，Es不但对裸鼠接种的膀胱癌有抑制作用，对膀胱癌细胞系也有抑制作用。Objective To explore the optimal method for protein expression in rhES （recombinant human endostatin ） and study the anti-tumor activities of rhES in solid tumor and established cell line. Methods Different IPTG concentrations, the timing of adding IPTG into the culture medium and the different time of expression were employed to explore the optimized conditions of protein expression. Activity examination： （1） animal experiment： nude mice bearing subcutaneous cancer in treated group and controlled group were observed. （2） cellular experiment： the inhibitory effect of rhES in T-24 established cell line were observed by MTT assay and cancer cell growth curve. Results The expression of rhES protein was 58.65%. Of all the E. eoli body proteins, the protein purity after purification was 96.22%. Activity examination indicated that rhES could inhibit the growth of solid tumor and the established cell line. In animal experiment, the tumor inhibition rate was 66.8%. Cell experiment： the 50% inhibitory concentration （IC50） was 22μg/ml. The cell population decreased 58.75% than the control group at Day 7 in the tumor cell growth curve. Conclusion A high expression and activity of rhES protein can be obtained by the optimized expression conditions, rhES can inhibit the cellular growth in both solid tumor and established cell line of bladder cancer.

关 键 词：内皮抑素类 膀胱肿瘤 蛋白表达 抗肿瘤 

分 类 号：R737.14[医药卫生—肿瘤]