Abstract | BACKGROUND: Micro- ribonucleic acid (miRNA)-199a-5p has been reported to be decreased in hepatocellular carcinoma (HCC) compared to normal tissue. Discoidin domain receptor-1 (DDR1) tyrosine kinase, involved in cell invasion-related signaling pathway, was predicted to be a potential target of miR-199a-5p by the use of miRNA target prediction algorithms. The aim of this study was to investigate the role of miR-199a-5p and DDR1 in HCC invasion. METHODS: Mature miR-199a-5p and DDR1 expression were evaluated in tumor and adjacent non- tumor liver tissues from 23 patients with HCC undergoing liver resection and five hepatoma cell lines by the use of real-time quantitative RT-PCR (qRT-PCR) analysis. The effect of aberrant miR-199a-5p expression on cell invasion was assessed in vitro using HepG2 and SNU-182 hepatoma cell lines. Luciferase reporter assay was employed to validate DDR1 as a putative miR-199a-5p target gene. Regulation of DDR1 expression by miR-199a-5p was assessed by the use qRT-PCR and western blotting analysis. RESULTS: A significant down-regulation of miR-199a-5p was observed in 65.2% of HCC tissues and in four of five cell lines. In contrast, DDR1 expression was significantly increased in 52.2% of HCC samples and in two of five cell lines. Increased DDR1 expression in HCC was associated with advanced tumor stage. DDR1 was shown to be a direct target of miR-199a-5p by luciferase reporter assay. Transfection of miR-199a-5p inhibited invasion of HepG2 but not SNU-182 hepatoma cells. CONCLUSIONS: Decreased expression of miR-199a-5p contributes to increased cell invasion by functional deregulation of DDR1 activity in HCC. However, the effect of miR-199a-5p on DDR1 varies among individuals and hepatoma cell lines. These findings may have significant translational relevance for development of new targeted therapies as well as prognostic prediction for patients with HCC.
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Authors | Qingli Shen, Vito R Cicinnati, Xiaoyong Zhang, Speranta Iacob, Frank Weber, Georgios C Sotiropoulos, Arnold Radtke, Mengji Lu, Andreas Paul, Guido Gerken, Susanne Beckebaum |
Journal | Molecular cancer
(Mol Cancer)
Vol. 9
Pg. 227
(Aug 27 2010)
ISSN: 1476-4598 [Electronic] England |
PMID | 20799954
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- MicroRNAs
- Receptors, Mitogen
- mirn199 microRNA, human
- Discoidin Domain Receptors
- Receptor Protein-Tyrosine Kinases
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Topics |
- Blotting, Western
- Carcinoma, Hepatocellular
(genetics, metabolism, pathology)
- Cell Line, Tumor
- Discoidin Domain Receptors
- Hep G2 Cells
- Humans
- Liver
(metabolism)
- Liver Neoplasms
(genetics, metabolism, pathology)
- MicroRNAs
(genetics, metabolism)
- Receptor Protein-Tyrosine Kinases
(genetics, metabolism)
- Receptors, Mitogen
(genetics, metabolism)
- Reverse Transcriptase Polymerase Chain Reaction
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