微小RNA-23a-p21活化激酶6通路对前列腺癌细胞迁移与侵袭能力的影响  

Impact of miRNA-23a-p21-activated kinase 6 signaling on the migration and invasion activities of prostate cancer calls

作　　者：蔡松旺[1] 李小娟[2] 陈锐涵[4] 朱宝益[3] 蔡燚[3] 叶春伟[3] 陶奕然[3] 冯淑君[5] 温星桥[3] 

机构地区：[1]中山大学附属第三医院心胸外科,广州510630 [2]中山大学附属第三医院保健科,广州510630 [3]中山大学附属第三医院泌尿外科,广州510630 [4]中山大学附属第三医院急诊外科,广州510630 [5]中山大学中山医学院

出　　处：《中华实验外科杂志》2012年第7期1360-1362,共3页Chinese Journal of Experimental Surgery

基　　金：基金项目：国家自然科学基金资助项目（81072115、30901768）;985创新团队项目（82000-3281901）;2011教育部新世纪人才支持项目;中央高校基本科研业务费专项资金资助项目（中山大学青年教师培育项目10ykpy06）

摘　　要：目的探讨调控p21活化激酶6（PAK6）的微小RNA（miRNA）与前列腺癌侵袭及迁移的关系。方法生物信息学预测靶向PAK6的miRNA，转染miRNA模拟物及抑制剂，Western blot法及实时定量逆转录-聚合酶链反应（Real—timePCR）检测PC-3细胞PAK6的表达，荧光素报告酶实验检测预测NmiRNA对PAK6的调控，并行体外迁移及侵袭实验，检测转染miRNA对PC-3细胞迁移及侵袭能力的影响。结果生物信息学预测miRNA-23a可能是调控PAK6的靶miRNA。Westernblot检测转染miRNA-23a组PAK6表达量下降79％，而转染miRNA-23a抑制剂组PAK6表达量上升40％，差异均有统计学意义（P〈0．05）。荧光素报告酶实验结果显示，转染miRNA-23a后野生型PAK6组荧光素酶活性下降32％，而突变组差异无统计学意义（P〉0．05）。Real-timePCR检测3组PAK6mRNA的表达，差异无统计学意义（P〉0．05）。体外迁移及侵袭实验示，转染miRNA-23a组迁移细胞数减少57％，侵袭的细胞数减少91％。结论微小RNA-23a通过下调靶蛋白PAK6的表达从而引起前列腺癌迁移及侵袭能力下降。Objective To investigate the role of the p21-activated kinase 6 （PAK6） and its target microRNA （miRNA） on prostate cancer cells migration and invasion. Methods MiRNA candidates which potentially target PAK6 was predicted by target prediction programs. The expression of PAK6 were meas- ured by real-time polymerase chain reaction （ Real-time PCR） and Western blotting after PC-3 cells were transfected with miR-23a mimics or inhibitor oligonucleotides. Luciferase reporter assay was used to deter- mine whether PAK6 was the direct target of miR-23a. Cell migration and invasion were detected by matrigel invasion assay and transwell migration assay. Results MiRNA 23a was identified by target prediction pro- grams. Exogenetic overexpression of miR-23a resulted in a remarkable decrease of PAK6 expression （79%）, whereas miR-23a inhibitor oligonucleotides induced pronounced increase of PAK6 expression （40%）. The luciferase activities were significantly inhibited 32% in wild-type PAK6 group （P 〈 0. 05 ）, while that were no significant difference in the mutation group （ P 〉 0.05 ）. PAK6 mRNA levels had no changed as detected by real-time PCR（ P 〉 0.05 ）. Matrigel invasion assay and transwell migration assay demonstrated that exogenetic overexpression of miR-23a markedly reduced migration and invasion of PC-3 cells （57%, 91% ）. Conclusion MiRNA-23a inhibited prostate cancer cell migration and invasion by repressing PAK6.

关 键 词：微小RNA PAK6 前列腺癌 转移 侵袭 

分 类 号：R737.25[医药卫生—肿瘤]