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JOURNAL ARTICLE
RESEARCH SUPPORT, NON-U.S. GOV'T
Identification of deregulated miRNAs and their targets in hepatitis B virus-associated hepatocellular carcinoma.
World Journal of Gastroenterology : WJG 2012 October 15
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.
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.
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