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Molecular Cancer
Published in: Molecular Cancer 1/2013

Open Access 01-12-2013 | Research

The identification of novel targets of miR-16 and characterization of their biological functions in cancer cells

Authors: Xin Yan, Hongwei Liang, Ting Deng, Kegan Zhu, Suyang Zhang, Nan Wang, Xueyuan Jiang, Xueliang Wang, Rui Liu, Ke Zen, Chen-Yu Zhang, Yi Ba, Xi Chen

Published in: Molecular Cancer | Issue 1/2013

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Abstract

Background

In eukaryotes, miR-16 is an important microRNA (miRNA) that is involved in numerous biological processes. However, it is not fully understood how miR-16 executes its physiological functions. In the present study, we aimed to identify novel miR-16 targets and study their biological functions.

Methods

Candidate target genes of miR-16 were screened by microarray analysis of mRNA levels in several cancer cell lines with enhanced miR-16. Three bioinformatics algorithms, including TargetScan, PicTar, and miRanda, were used in combination to calculate the miR-16 targets. The expression levels of miR-16 and target mRNA were examined by relative quantification RT-PCR, and the expression levels of target protein were detected by Western blot. Luciferase reporter plasmids were constructed to confirm direct targeting. The effect of miR-16 and target gene on cell viability was evaluated using MTT assays. The effects of miR-16 and target gene on apoptosis and cell cycle distribution were evaluated by flow cytometry analysis.

Results

By overexpressing miR-16 in several cancer cell lines and measuring global mRNA levels using microarray analysis, we identified 27 genes that may be regulated by miR-16. After the bioinformatics filtering process, 18 genes were selected as candidate miR-16 targets. Furthermore, we experimentally validated three of these candidates, MAP7 (microtubule-associated protein 7), PRDM4 (PR domain containing 4) and CDS2 (CDP-diacylglycerol synthase 2), as direct targets of miR-16. Finally, we demonstrated that miR-16 targeting MAP7 played a critical role in regulating proliferation but not apoptosis and cell cycle progression in cancer cells.

Conclusion

In summary, the present study identifies several novel miR-16 targets and illustrates a novel function of miR-16 targeting MAP7 in modulating proliferation in cancer cells.
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Metadata
Title
The identification of novel targets of miR-16 and characterization of their biological functions in cancer cells
Authors
Xin Yan
Hongwei Liang
Ting Deng
Kegan Zhu
Suyang Zhang
Nan Wang
Xueyuan Jiang
Xueliang Wang
Rui Liu
Ke Zen
Chen-Yu Zhang
Yi Ba
Xi Chen
Publication date
01-12-2013
Publisher
BioMed Central
Published in
Molecular Cancer / Issue 1/2013
Electronic ISSN: 1476-4598
DOI
https://doi.org/10.1186/1476-4598-12-92

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