CAJ | 학술논문

目的探讨微RNA(miRNA)-21在肝细胞癌中的表达特征及其功能,筛选并鉴定miRNA-21可能调控的靶基因.方法用PAGE/Northern blot方法分析miRNA-21在10例肝细胞癌患者组织标本中的表达特征,并在3种肝癌细胞株中加以验证.用生物信息学分析预测miRNA-21可能的靶基因,通过逆转录-聚合酶链反应(RT-PCR)方法克隆靶基因的调控区序列,利用荧光素酶双报告基因系统筛选,并通过Western blot、流式细胞仪在蛋白质水平进行分析验证.用体外细胞侵袭实验分析miRNA-21与肝癌细胞侵袭能力的关系.结果 miRNA-21在10例肝细胞癌患者中均表达上调,与癌旁组织比较,表达量增加10倍以上.在3种肝癌细胞株中也得到相似的结果.筛选并确认了RECK为miRNA-21的调控靶基因之一.抑制miRNA-21的表达可明显降低肝癌细胞的侵袭.结论 miRNA-21在肝细胞癌中表达失控,通过抑制RECK表达促进肿瘤细胞侵袭.
목적 탐토미RNA(miRNA)-21재간세포암중적표체특정급기공능,사선병감정miRNA-21가능조공적파기인.방법 용PAGE/Northern blot방법분석miRNA-21재10례간세포암환자조직표본중적표체특정,병재3충간암세포주중가이험증.용생물신식학분석예측miRNA-21가능적파기인,통과역전록-취합매련반응(RT-PCR)방법극륭파기인적조공구서렬,이용형광소매쌍보고기인계통사선,병통과Western blot、류식세포의재단백질수평진행분석험증.용체외세포침습실험분석miRNA-21여간암세포침습능력적관계.결과 miRNA-21재10례간세포암환자중균표체상조,여암방조직비교,표체량증가10배이상.재3충간암세포주중야득도상사적결과.사선병학인료RECK위miRNA-21적조공파기인지일.억제miRNA-21적표체가명현강저간암세포적침습.결론 miRNA-21재간세포암중표체실공,통과억제RECK표체촉진종류세포침습.
Objective To research the expression signature and function of miRNA-21 in hepatocellular carcinoma (HCC), and indentify its functional target mRNA.Methods The expression signature of miRNA-21 in 10 cases of HCC was confirmed in 3 kinds of HCC cell lines by PAGE/Northern blot.The potential target mRNA prediction of miRNA-21 was performed by bioinformatics.The regnlation region of miRNA-21 in RECK mRNA was cloned by RT-PCR, and confirmed by sequencing.Dual report gene assay was used to identify the target gene of miRNA-21.The target gene was confirmed by Western blot and FCM analysis.The effect of miRNA-21 on HCC cell line HepG2 was analyzed by in vitro invasion assay.Results miRNA-21 was over-expressed in 10 cases of HCC and 3 kinds of cell lines.RECK was identified as one of miRNA-21 target genes.The invasion ability of HepG2 was suppressed when miRNA-21 was knocked down.Conclusion miRNA-21 was over-expressed in HCC, and promoted cancer cell invasion by inhibiting the expression level of RECK.