PTEN and PDCD4 are Bona Fide Targets of microRNA-21 in Human Cholangiocarcinoma  

作　　者：Chang-zheng Liu Wei Liu Yi Zheng Jin-mei Su Jing--jing Li Lan Yu Xiao-dong He Song-sen Chen 

机构地区：[1]Department of Biochemistry, National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China [2]Department of General Surgery [3]Department of Rheumatology,Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, 100730, China

出　　处：《Chinese Medical Sciences Journal》2012年第2期65-72,共8页中国医学科学杂志（英文版）

基　　金：Supported by Institute of Basic Medical Sciences(2009PY13 and 2010PYZ18);the National Natural Science Foundation of China(81100608 and 30901342)

摘　　要：Objective To investigate the expression profile of microRNA-21 in human cholangiocarcinoma tis- sues and to validate its bona fide targets in human cholangiocarcinoma cells. Methods The expression profile of microRNA-21 in human cholangiocarcinoma tissues and cholan- giocarcinoma cell line, QBC939, was evaluated by using real-time PCR analysis. The bona fide targets of microRNA-21 were analyzed and confirmed by dual luciferase reporter gene assay and western blot, respec- tively. The expressional correlation of microRNA-21 and its targets was probed in human cholangiocarci- noma tissues by using real-time PCR, locked nucleic acid in situ hybridization （LNA-ISH）, and immunohis- tochemistry analysis. Results Real-time PCR analysis revealed that microRNA-21 expression depicted a significant up-regulation in human cholangiocarcinoma tissues about 5.6-fold as compared to the matched normal bileduct tissues （P〈0.05）. The dual luciferase reporter gene assay revealed endogenous microRNA-21 in cholan- giocarcinoma cell line, QBC939, inhibited the luciferase reporter activities of wild-type PTEN （P〈0.01） and PDCD4 （P〈0.05） and had no this effect on mutated PTEN and PDCD4. Moreover, loss of microRNA-21 function led to a significant increase of PTEN and PDCD4 protein levels in QBC939 cells. Elevated microRNA-21 levels were accompanied by marked reductions of PTEN and PDCD4 expression in the same cholangiocarcinoma tissue. Conclusion microRNA-21 expression is up PDCD4 are direct effectors of microRNA-21. regulated in human cholangiocarcinoma and PTEN,

关 键 词：CHOLANGIOCARCINOMA MICRORNA-21 phosphatase and tensin homolog programmed cell death 4 

分 类 号：Q786[生物学—分子生物学] Q279