Brief Article
Copyright ©2012 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Oct 14, 2012; 18(38): 5442-5453
Published online Oct 14, 2012. doi: 10.3748/wjg.v18.i38.5442
Identification of deregulated miRNAs and their targets in hepatitis B virus-associated hepatocellular carcinoma
Wen Wang, Lan Juan Zhao, Ye-Xiong Tan, Hao Ren, Zhong-Tian Qi
Wen Wang, Hao Ren, Zhong-Tian Qi, Lan-Juan Zhao, Department of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Second Military Medical University, Shanghai 200433, China
Ye-Xiong Tan, Laboratory of Signal Transduction, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai 200433, China
Author contributions: Wang W and Zhao LJ contributed equally to the work, Wang W performed the research, drafted the article; Zhao LJ performed the research and revised the article; Tan YX performed the research; Ren H and Qi ZT designed the research; and all the authors have read and approved the final version to be published.
Supported by The Key Programs of the Ministry of Science and Technology, No. 2012ZX10002009-004; Shanghai Leading Academic Discipline Project (B901) and Science Fund for Creative Research Groups, NSFC, China, No. 30921006
Correspondence to: Zhong-Tian Qi, MD, PhD, Department of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Second Military Medical University, 800 Xiangyin Road, Shanghai 200433, China. qizt@smmu.edu.cn
Telephone: +86-21-81870988 Fax: +86-21-81870988
Received: January 16, 2012
Revised: March 28, 2012
Accepted: April 9, 2012
Published online: October 14, 2012
Abstract

AIM: To identify the differentially expressed miRNAs and their targets in hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC).

METHODS: Six hundred and sixty seven human miRNAs were quantitatively analyzed by Taqman low-density miRNA array (TLDA) in HBV-HCC tissues. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were used to analyze the significant function and pathway of the differentially expressed miRNAs in HBV-HCC. TargetScan software was used to predict the targets of deregulated miRNAs. Western blotting and luciferase assay were performed to verify the targets of these miRNAs.

RESULTS: Ten up-regulated miRNAs (miR-217, miR-518b, miR-517c, miR-520g, miR-519a, miR-522, miR-518e, miR-525-3p, miR-512-3p, and miR-518a-3p) and 11 down-regulated miRNAs (miR-138, miR-214, miR-214#, miR-199a-5p, miR-433, miR-511, miR-592, miR-483-3p, miR-483-5p, miR-708 and miR-1275) were identified by Taqman miRNAs array and confirmed quantitatively by reverse transcription polymerase chain reaction in HCC and adjacent non-tumor tissues. GO and KEGG pathway analysis revealed that “regulation of actin cytoskeleton” and “pathway in cancer” are most likely to play critical roles in HCC tumorigenesis. MiR-519a and ribosomal protein S6 kinase polypeptide 3 (RPS6KA3) were predicted as the most significant candidates by miRNA-mRNA network. In addition, cyclin D3 (CCND3) and clathrin heavy chain (CHC), usually up-regulated in HCC tissues, were validated as the direct target of miR-138 and miR-199a-5p, respectively.

CONCLUSION: Our data suggest an importance of miR-138 and miR-199a-5p as well as their targets CCND3 and CHC in HCC tumorigenesis, and may provide more evidence for reliability of integrative bioinformatics analysis.

Keywords: Hepatocellular carcinoma, miR-138, miR-199a-5p, Cyclin D3, Clathrin heavy chain, Bioinformatics, Taqman array