Prediction and identification of microRNAs involved in regulating neuropilin-2 in colorectal carcinoma

Aim: To predict and identify the possible microRNAs (miRNAs) involved in regulating neuropilin- 2 (NRP2) in colorectal carcinoma. Methods: Bioinformatic technique was used to predict the possible miRNAs which might participate in regulating NRP2. miR-486-5p, which is most possibly involved in regulating NRP2, was selected for further study. A miR- 486-5p over-expressing plasmid was constructed and transfected into the human colorectal carcinoma cell line SW620 using Lipofectamine 2000. The expression of miR-486-5p in transfected cells was measured by quantitative real-time PCR (qRT-PCR). Western blot was conducted to detect the expression of NRP2 protein, and miR- 486-5p targeting NRP2 3'UTR was verified by dual luciferase reporter assay. Results: Twenty-five possible NRP2-regulating miRNAs were predicted using bioinformatic software. By searching the literature, we chose miR-486-5p for further study. The expression of miR-486-5p in SW620 cells transfected with miR- 486-5p over-expressing plasmid was obviously up-regulated. Western blot analysis revealed that miR-486-5p reduced the expression level of NRP2. Dual luciferase reporter assay demonstrated that miR-486-5p could directly target the NRP2 3'UTR. Conclusion: miR-486-5p is involved in the post-transcriptional regulation of NRP2 by binding to the NRP2 3'UTR and interfering the normal translation of NRP2. ? 2014 Baishideng Publishing Group Co., Limited. Allrights reserved.