基础医学与临床 ›› 2012, Vol. 32 ›› Issue (6): 628-633.

• 研究论文 • 上一篇    下一篇

利用microRNA-基因芯片联合Real-Time PCR方法鉴定肝内及肝外胆管癌表达下调的microRNAs

刘长征1,何小东2,于岚3,刘卫3,陈松森1,李静静4,蔡磊5   

  1. 1. 中国医学科学院基础医学研究所
    2. 北京协和医院
    3. 中国医学科学院北京协和医院
    4. 中国医学科学院北京协和医院基本外科
    5. 中国医学科学院整形外科医院
  • 收稿日期:2012-03-15 修回日期:2012-03-22 出版日期:2012-06-05 发布日期:2012-05-25
  • 通讯作者: 陈松森 E-mail:chensongsen@ibms.pumc.edu.cn
  • 基金资助:
    重塑FOXA2-miR-29反馈调控网络可能恢复糖尿病小鼠血糖的稳定调节

Identification of down-regulated miRNAs in extrahepatic and intrahepatic cholangiocarcinoma by using microRNA-microarray and Real-Time PCR analysis.

  • Received:2012-03-15 Revised:2012-03-22 Online:2012-06-05 Published:2012-05-25

摘要: 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进行初步的功能研究,即利用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,肝外胆管癌组织表达特异表达下调的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中过表达分别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。

关键词: 胆管癌, microRNA, 细胞增殖

Abstract: Objective We intend to investigate the expression profile of miRNAs down-regualted in human extrahepatic and intrahepatic cholangiocarcinoma tissues and 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 using miRNA-microarray, which was confirmed by using 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. Conclusion The miRNAs expression patterns in human extrahepatic and intrahepatic cholangiocarcinoma tissues are different and Down-regulated miRNAs act as suppressors on cholangiocarcinoma cell growth.

Key words: cholangiocarcinoma, microRNAs, cell growth