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Archive > Volume 21 Issue 3 > 2014,21(3):298-302. DOI:10.3872/j.issn.1007-385X.2014.03.011 Prev Next
Basic Research
Effect of miRNA-486-5p on expression of NRP2 in gastric carcinoma SGC7901 cells
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Abstract:
To predict and identify the target genes of microRNA-486-5p (miRNA-486-5p) in human gastric cancer SGC7901 cells. Methods: Possible target genes of miRNA-486-5p were predicted by bioinformatics techniques and accordingly miRNA-486-5p over-expressing plasmid (GV214-miR) against the identified target gene, neuropilin-2 (NRP-2) was constructed. SGC7901 cells were transfected with a control miRNA and an NRP-2-specific miRNA-486-5p. In the transfectants and non-transfected control cells, miRNA-486-5p and NRP-2 mRNA levels and NRP-2 protein levels were analyzed by real-time PCR and Western blotting respectively, and the NRP-2 promoter activity was evaluated by a dual luciferase reporter assay. Results: The expression of miRNA-486-5p in miRNA-486-5p-transfected SGC7901 cells (SGC7901-miR cells) was significantly up-regulated compared with that in the control group (8.21±118 vs 1.02±026, P<0.01). No significant difference in NRP2 mRNA abundance was observed (P>0.05). However, the NRP2 protein level was significantly reduced in SGC7901-miR cells (0.36±0.06) as compared with SGC7901 cells transfected with the control plasmid (0.76±0.05, P<0.05). Dual luciferase reporter assay demonstrated that miRNA-486-5p directly targeted the 3’-untranslated region (UTR) of the NRP2 gene, resulting in inhibition of the post-transcriptional translation of NRP2. Conclusion: Sequence-specific miRNA-486-5p may suppress the expression of NRP2 at the protein level in human gastric cancer cells by binding to NRP2 mRNA 3’UTR directly.
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Project Supported:
Project supported by Higher Educational Science and Technology Foundation of Shandong Province (No. J11LF75), and the Scientific Research Foundation of Binzhou Medical College (No. BY2010KYQD02)
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