探寻肝再生磷酸酶3在膀胱癌细胞T24中的作用  

Exploring the Function of Phosphatase of Regenerating Liver-3 in Bladder Cancer cell Line-T24

作　　者：薛庆[1] 于垂恭[1] 于磊[1] 武国军[1] 袁建林[1] 

机构地区：[1]第四军医大学西京医院泌尿外科,陕西西安710032

出　　处：《现代生物医学进展》2013年第25期4829-4832,4850,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(30872583)

摘　　要：目的:已经有研究证明肝再生磷酸酶3(Phosphatase of Regenerating Liver-3,PRL-3)在消化道肿瘤的发生发展过程中起到重要作用,但其在膀胱癌中发挥何种作用尚不清楚,本次研究主要探寻PRL-3在膀胱癌细胞T24中作用,以求为膀胱癌的治疗提供新思路。方法:采用siRNA干涉的方法下调T24中PRL-3蛋白表达水平,通过western blot方法检测干涉效果,绘制细胞生长曲线、构建裸鼠种植瘤模型,检测PRL-3下调对细胞体外及体内增殖的影响。结果:我们所设计的siRNA能够下调PRL-3在A498细胞中的表达(F=7.26,P<0.05),细胞生长曲线显示下调PRL-3能够抑制细胞的增殖,这种作用从干涉后的第60h开始,随时间增加作用愈加明显(F≥8.35,P<0.05);动物实验表明,siRNA下调PRL-3能够抑制T24细胞在活体内的增殖,从干涉后第21天开始这种作用开始体现出来(F≥10.46,P<0.05),并且干涉组的肿瘤肿瘤明显低于两个对照组(F=13.63,P<0.05)。结论:我们构建的siRNA能够在T24细胞中实现对PRL-3蛋白的下调,siRNA介导的PRL-3蛋白下调能够明显抑制T24细胞在活体外及活体内的增殖,这初步证明PRL-3在膀胱癌中发挥重要的作用。随着我们对PRL-3分子进一步深入的了解,揭示其发挥作用的具体机制,PRL-3很可能成为膀胱癌基因治疗的新靶点。Objoetive： Other previous reseachers have verified that phosphatase of regenerating livere-3 （PRL-3） played a crucial role in digestive -tract cancer, but its function in bladder cancer is unknown. This research explored the function of PRL-3 in bladder can- cer cell line-T24 for finding new ideas in bladder cancer therapy. Methods： PRL-3 siRNA was designed and transfected into T24 cell （the tmnsfection was performed using Lipofectamine TM 2000）. The interfering effects were detected on protein levels using western blot. The affection of the siRNA on cell proliferation was observed through cell growth curves in vitro, tumor growth curves in vivo. Results： The results of western blot indicated that the siRNA we designed could down-regulate the expression of PRL-3 in T24 cell properly （F=7. 26 ,P〈0.05）. Cell growth curves indicated that down-regulation of PRL-3 could suppress the proliferation of the T24 cells in vitro, and the suppression occurred at 60h after interference and strengthen with time from then （F≥ 8.35 ,P〈0.05）. The tumor growth curves showed that down-regulation of PRL-3 could suppress the proliferation of the T24 cells on 21d after interference in vivo （F〉 10.46, P〈0.05）, and the tumor weights in interfering group were lower than those of control groups （F=13.63, P〈0.05）. Conclusion： The siRNA that we con- strutted could inhibit the expression of PRL-3 protein in T24 cell, Down-regulation of PRL-3 mediated by siRNA could suppress the pro- liferation of the T24 cells significantly both in vitro and in vivo. These results initially proved that PRL-3 played a crucial role in bladder cancer. With the development of PRL-3 researches, the mechanism of PRL-3 achieving its function would be revealed. And PRL-3 might become a new target gene for bladder cancer therapy.

关 键 词：膀胱癌 PRL-3 SIRNA 基因治疗 

分 类 号：R737.14[医药卫生—肿瘤]