Ren Ming-hua, Ni Shao-bin, Chen Qi-yin, Xu Wan-hai, Wang Chang-lin, Fu Yi-ming, Jiao Zhi-xing, Ma Li, Zhao Zhong-shan, Liu Xing-han
Department of Urology, First Affiliated Hospital of Harbin Medical University, Harbin 150001, China.
Zhonghua Yi Xue Za Zhi. 2009 Dec 15;89(46):3285-8.
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.
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.
The expression of rhES protein was 58.65%. Of all the E. coli 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 (IC(50)) was 22 microg/ml. The cell population decreased 58.75% than the control group at Day 7 in the tumor cell growth curve.
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.
探索重组人内皮抑素（rhES）蛋白表达的最佳方法，并研究rhES在实体瘤及已建立的细胞系中的抗肿瘤活性。
采用不同的异丙基-β-D-硫代半乳糖苷（IPTG）浓度、向培养基中添加IPTG的时间以及不同的表达时间来探索蛋白表达的优化条件。活性检测：（1）动物实验：观察治疗组和对照组皮下荷瘤裸鼠。（2）细胞实验：通过MTT法和癌细胞生长曲线观察rhES对已建立的T-24细胞系的抑制作用。
rhES蛋白表达量为58.65%。在所有大肠杆菌菌体蛋白中，纯化后蛋白纯度为96.22%。活性检测表明，rhES可抑制实体瘤和已建立的细胞系的生长。动物实验中，肿瘤抑制率为66.8%。细胞实验：50%抑制浓度（IC50）为22μg/ml。在肿瘤细胞生长曲线中，第7天时细胞数量比对照组减少58.75%。
通过优化表达条件可获得高表达且具有活性的rhES蛋白。rhES可抑制实体瘤和膀胱癌已建立细胞系中的细胞生长。
