肝内及肝外胆管癌表达下调microRNAs表达谱的鉴定

目的筛选并鉴定肝内及肝外胆管癌组织表达下调的miRNAs,进行初步的功能研究。方法利用miRNA-基因芯片方法筛选肝内、肝外胆管癌组织特异表达下调或共同表达下调的miRNAs,选择real-time PCR方法进行验证;根据肝内及肝外胆管癌不同的下调miRNAs表达谱,选择差异明显且具有代表性的miRNAs,利用miRNA特异模拟物转染胆管癌细胞系QBC939,利用MTT试剂盒分析细胞增殖变化,研究表达下调miRNAs与胆管癌细胞增殖的关系。结果表达分析显示,肝外胆管癌组织特异表达下调的miRNAs共25个,肝内胆管癌组织特异表达下调的miRNAs共15个,在肝内及肝外胆管癌组织均表达下调的miRNAs共6个。在胆管癌细胞系QBC939中分别过表达miR-181a、miR-596、miR-492以及miR-602可以明显抑制细胞增殖。结论肝内及肝外胆管癌具有不同的miRNAs表达谱,表达下调的miRNAs可以明显抑制胆管癌细胞增殖。

Identification of down-regulated miRNAs in extrahepatic and intrahepatic cholangiocarcinoma

Objective To investigate the expression profile of miRNAs down-regualted in human extrahepatic and intrahepatic cholangiocarcinoma tissues and to probe the effect on cell growth of four of these miRNAs in QBC939 cell line.Methods Down-regulated miRNAs in extrahepatic or intrahepatic cholangiocarcinoma tissues were analyzed by miRNA-microarray and confirmed by miRNA real-time PCR analysis.Based on these findings,four of these down-regulated miNRAs were chosen to perform function investigation.The miRNAs mimics were transfected into QBC939 cells and cell proliferation assay was performed by using MTT,which would demonstrate the relationship between down-regulated miRNAs and cholangiocarcinoma cell growth.Results 25 miRNAs and 15 miRNAs were down-regulated in extrahepatic cholangiocarcinoma and intrahepatic cholangiocarcinoma,respectively.There were 6 miRNAs down-regulated both in two types of cholangiocarcinoma.miR-181a,miR-596,miR-492,and miR-602 were overexpressed in QBC939 cells,which indicated a significant inhibitory effect on cell growth. Conclusions The miRNAs expression patterns in human extrahepatic and intrahepatic cholangiocarcinoma tissues are different and down-regulated miRNAs act as suppressors on cholangiocarcinoma cell growth.