CAJ | 학술논문

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-time PCR)检测PC-3细胞PAK6的表达,荧光素报告酶实验检测预测miRNA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. Western blot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-time PCR检测3组PAK6 mRNA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.
목적 탐토조공p21활화격매6(PAK6)적미소RNA(miRNA)여전렬선암침습급천이적관계.방법 생물신식학예측파향PAK6적miRNA,전염miRNA모의물급억제제,Western blot 법급실시정량역전록-취합매련반응( Real-time PCR)검측PC-3세포PAK6적표체,형광소보고매실험검측예측miRNA대PAK6적조공,병행체외천이급침습실험,검측전염miRNA대PC-3세포천이급침습능력적영향.결과 생물신식학예측miRNA-23a가능시조공PAK6적파miRNA. Western blot검측전염miRNA-23a조PAK6표체량하강79％,이전염miRNA-23a억제제조PAK6표체량상승40％,차이균유통계학의의(P＜0.05).형광소보고매실험결과현시,전염miRNA-23a후야생형PAK6조형광소매활성하강32％,이돌변조차이무통계학의의(P＞0.05).Real-time PCR검측3조PAK6 mRNA적표체,차이무통계학의의(P＞0.05).체외천이급침습실험시,전염miRNA-23a조천이세포수감소57％,침습적세포수감소91％.결론 미소RNA-23a통과하조파단백PAK6적표체종이인기전렬선암천이급침습능력하강.
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 measured 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 determine whether PAK6 was the direct target of miR-23a.Cell migration and invasion were detected hy matrigel invasion assay and transwell migration assay.Results MiRNA 23a was identified by target prediction programs. 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.