MicroRNA-510 promotes cell and tumor growth by targeting peroxiredoxin1 in breast cancer

Breast Cancer Research

Victoria J Findlay²  David P Turner²  Amanda C Larue²  E Ramsay Camp¹  Natalie J Mason²  Lourdes M Nogueira²  Savannah G Bandurraga²  Jamie N Mills²  Qi J Guo² 

[1] Department of Surgery, 25 Courtenay Drive, Medical University of South Carolina, Charleston, SC 29425, USA;Department of Pathology & Laboratory Medicine, 39 Sabin Street, Medical University of South Carolina, Charleston, SC 29425, USA

关键词: miR-510;    migration;    breast cancer;    tumorigenesis;    peroxiredoxin1;    MicroRNA;   

Others  :  793760
DOI  :  10.1186/bcr3464

 received in 2012-09-23, accepted in 2013-08-23,  发布年份 2013

【 摘 要 】

Introduction

MicroRNAs are small non-coding RNAs that are involved in the post-transcriptional negative regulation of mRNAs. MicroRNA 510 (miR-510) was initially shown to have a potential oncogenic role in breast cancer by the observation of its elevated levels in human breast tumor samples when compared to matched non-tumor samples. Few targets have been identified for miR-510. However, as microRNAs function through the negative regulation of their direct targets, the identification of those targets is critical for the understanding of their functional role in breast cancer.

Methods

Breast cancer cell lines were transfected with pre-miR-510 or antisense miR-510 and western blotting and quantitative real time PCR were performed. Functional assays performed included cell growth, migration, invasion, colony formation, cytotoxicity and in vivo tumor growth. We performed a PCR assay to identify novel direct targets of miR-510. The study focused on peroxiredoxin 1 (PRDX1) as it was identified through our screen and was bioinformatically predicted to contain a miR-510 seed site in its 3' untranslated region (3'UTR). Luciferase reporter assays and site-directed mutagenesis were performed to confirm PRDX1 as a direct target. The Student's two-sided, paired t-test was used and a P-value less than 0.05 was considered significant.

Results

We show that miR-510 overexpression in non-transformed and breast cancer cells can increase their cell growth, migration, invasion and colony formation in vitro. We also observed increased tumor growth when miR-510 was overexpressed in vivo. We identified PRDX1 through a novel PCR screen and confirmed it as a direct target using luciferase reporter assays. The reintroduction of PRDX1 into breast cancer cell lines without its regulatory 3'UTR confirmed that miR-510 was mediating its migratory phenotype at least in part through the negative regulation of PRDX1. Furthermore, the PI3K/Akt pathway was identified as a positive regulator of miR-510 both in vitro and in vivo.

Conclusions

In this study, we provide evidence to support a role for miR-510 as a novel oncomir. We show that miR-510 directly binds to the 3'UTR of PRDX1 and blocks its protein expression, thereby suppressing migration of human breast cancer cells. Taken together, these data support a pivotal role for miR-510 in breast cancer progression and suggest it as a potential therapeutic target in breast cancer patients.

【 授权许可】

   
2013 Guo et al.; licensee BioMed Central Ltd.

【 图 表 】

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