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

Molecular Cancer

Research

Hongwei Liang¹  Xueliang Wang¹  Suyang Zhang¹  Xueyuan Jiang¹  Xi Chen¹  Chen-Yu Zhang¹  Kegan Zhu¹  Nan Wang¹  Ke Zen¹  Ting Deng²  Rui Liu²  Yi Ba²  Xin Yan² 

[1] Jiangsu Engineering Research Center for microRNA Biology and Biotechnology, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, 22 Hankou Road, 210093, Nanjing, China;Tianjin Medical University Cancer Institute and Hospital, Huanhuxi Road, 300060, Tiyuanbei, Tianjin, China;

关键词: microRNA;    miR-16;    MAP7;    PRDM4;    CDS2;   

DOI  :  10.1186/1476-4598-12-92

 received in 2013-02-07, accepted in 2013-08-13,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundIn 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.MethodsCandidate 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.ResultsBy 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.ConclusionIn 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.

【 授权许可】

Unknown   
© Yan et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.