Visualization and quantitation of the expression of microRNAs and their target genes in neuroblastoma single cells using imaging cytometry

BMC Research Notes

Natasha S Barteneva¹  Tatiana Veremeyko²  Eugene D Ponomarev² 

[1]Department of Pediatrics, Harvard Medical School, Boston, MA, USA

[2]Center for Neurologic Diseases, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA

关键词: CDK6;    MiR-124;    Neuroblastoma;    Imaging cytometry;    Target gene;    MicroRNA;   

Others  :  1166914
DOI  :  10.1186/1756-0500-4-517

 received in 2011-08-15, accepted in 2011-11-28,  发布年份 2011

【 摘 要 】

Background

MicroRNAs (miRNAs) are regulatory molecules that play an important role in many physiological processes, including cell growth, differentiation, and apoptosis. In addition to modulating normal cellular functions, it has also been reported that miRNAs are involved in the development of many pathologies, including cardiovascular diseases, cancer, inflammation, and neurodegeneration. Methods for the sensitive detection and measurement of specific miRNAs and their cellular targets are essential for both basic research endeavours, as well as diagnostic efforts aimed at understanding the role of miRNAs in disease processes.

Findings

In this study, we describe a novel, imaging cytometry-based protocol that allows for simultaneous visualisation and quantification of miRNAs and their putative targets. We validated this methodology in a neuronal cell line by examining the relationship of the miRNA miR-124 and its known target, cyclin dependent kinase 6 (CDK6). We found that ectopic overexpression of miR-124 resulted in the downregulation of CDK6, decreased cellular proliferation, and induced cellular morphological changes.

Conclusions

This method is suitable for analysing the expression and cellular localisation of miRNAs and target proteins in small cell subsets within a heterogeneous cell suspension. We believe that our cytometry-based methodology will be easily adaptable to miRNA studies in many areas of biomedical research including neuroscience, stem cell biology, immunology, and oncology.

【 授权许可】

   
2011 Ponomarev et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

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