RNA激活技术介导的E-cadherin基因上调表达抑制5637膀胱癌细胞侵袭与迁移  

Up-regulation of E-cadherin by small activating RNA inhibits cell invasion and migration in 5637 human bladder cancer cells

作　　者：毛祺琦[1] 谢立平[1] 郑祥毅[1] 杨凯[1] 秦杰[1] 白宇[1] 王云彬[1] 

机构地区：[1]浙江大学医学院附属第一医院泌尿外科,杭州310003

出　　处：《现代泌尿生殖肿瘤杂志》2009年第1期34-38,共5页Journal of Contemporary Urologic and Reproductive Oncology

基　　金：国家自然科学基金资助项目(30770832;30801370);浙江省自然科学基金资助项目(Y207415)

摘　　要：目的探讨通过RNA激活(RNAactivation,RNAa)技术上调E-cadherin基因的表达而影响人膀胱癌5637细胞的侵袭、迁移能力。方法将与E-cadherin基因启动子DNA序列互补的双链RNA分子(dsEcad)转染入5637细胞中,采用RT-PCR法及Western blot检测E-cadherin的表达;用Transwell小室法及划痕法检测RNAa后细胞侵袭、迁移能力的改变;用细胞免疫荧光法及Western blot检测β-catenin蛋白在细胞内的重新分布结果。结果dsEcad转染5637细胞72 h后E-cadherin表达显著上调,RNAa有效;5637细胞对细胞外基质的侵袭能力及迁移能力也明显下降。β-catenin在胞核及细胞质的水平明显下降并重新再分布至细胞膜上。结论RNAa技术激活E-cadherin基因表达并抑制细胞侵袭、转移等恶性行为,可作为膀胱癌或其他恶性肿瘤基因治疗的一种有效手段。Objective To observe the effects of E-cadherin activation by RNAa on bladder cancer cell migration and invasion. Methods A dsRNA （dsEcad） targeting the E-cadherin gene promoter at position-215 relative to the transcription start site was transfected into 5637 cells. Expression of mRNA and protein was evaluated by semiquantitative RT-PCR and Western blot. Matrigel invasion assays and wound closure assays were performed to examine the migrative and invasive ability of the 5637 cells. Imrnunofluorescence was performed to demonstrate the relocalization of β- catenin. Results dsEcad caused a greater than 2-fold induction in E-cadherin expression and inhibit migration and invasion of 5637 bladder cancer cells in vitro, this inhibitory effect was associated with relocalization of β-catenin from the nucleus to the plasma membrane and decreased β-catenin-mediated transactivation. Conclusions Activation of E-cadherin by saRNA may have a therapeutic benefit for bladder and other types of cancer.

关 键 词：膀胱肿瘤 RNA激活技术 E-CADHERIN 侵袭 

分 类 号：R737.14[医药卫生—肿瘤]