title: Impact of miR-7 over-expression on the proteome of Chinese hamster ovary cells.
abstract: MicroRNAs play critical roles in the regulation of biological processes such as growth, apoptosis, productivity and secretion thus representing a potential route toward enhancing desirable characteristics of mammalian cells for biopharmaceutical production. We have previously found that miR-7 over-expression significantly inhibits the growth of CHO-SEAP cells without impacting cellular viability, with an associated increase in normalised productivity. Understanding the biological basis of this effect might open the way to new strategies for bioprocess-relevant growth regulation. In this study we have carried out a quantitative label-free LC-MS profiling study of proteins exhibiting altered levels following over-expression of miR-7 to gain insights into potential mechanisms involved in the observed phenotype. From the analysis we found 93 proteins showing decreased levels and 74 proteins with increased levels following over-expression of miR-7. Pathway analysis suggests that proteins involved in protein translation (e.g. ribosomal proteins), RNA and DNA processing (including histones) are enriched in the list of proteins showing decreased expression. Proteins involved in protein folding and secretion were found to be up-regulated following miR-7 over-expression. In silico bioinformatic analysis using miRWalk, which combined the output from 6 selected miRNA target prediction algorithms, was used to evaluate if any of the down-regulated proteins were potential direct targets of miR-7. Two genes, stathmin and catalase, which both have known roles in the regulation of cellular growth, were found to overlap a number of the predictive target database searches in both mouse and rat, and are likely to be possible direct targets of miR-7 in CHO cells. This is the first report investigating the impact of a miRNA on the proteome of CHO cells.
authors.affiliation: National Institute for Cellular Biotechnology, Dublin City University, Glasnevin, Dublin 9, Ireland. Paula.Meleady@dcu.ie
authors.forename: Paula
authors.initials: P
authors.lastname: Meleady
authors.forename: Mark
authors.initials: M
authors.lastname: Gallagher
authors.forename: Colin
authors.initials: C
authors.lastname: Clarke
authors.forename: Michael
authors.initials: M
authors.lastname: Henry
authors.forename: Noelia
authors.initials: N
authors.lastname: Sanchez
authors.forename: Niall
authors.initials: N
authors.lastname: Barron
authors.forename: Martin
authors.initials: M
authors.lastname: Clynes
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D020543:Proteome / Q000032:analysis* / Q000378:metabolism*; D015854:Up-Regulation / Q000502:physiology*
chemical_list: C575438:MIRN71 microRNA, C elegans; D035683:MicroRNAs; D020543:Proteome