Microarray and proteomics expression profiling identifies several candidates, including the valosin-containing protein (VCP), involved in regulating high cellular growth rate in production CHO cell lines
| Autor: | Niall Barron, Martin S. Sinacore, Kevin M. McCarthy, Padraig Doolan, Michael Henry, Martin Clynes, Ross Gallagher, Patrick Gammell, Paula Meleady, Mark Melville, Timothy S. Charlebois, Mark Leonard |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Proteome
Gene Expression Bioengineering Cell Cycle Proteins CHO Cells Biology Applied Microbiology and Biotechnology Cricetulus Valosin Containing Protein Cricetinae Gene expression Animals Gene Silencing RNA Small Interfering Cell Cycle Protein Adenosine Triphosphatases Gene knockdown Cell growth Chinese hamster ovary cell Gene Expression Profiling Cell Cycle Epithelial Cells Cell cycle Molecular biology Up-Regulation Gene expression profiling Cell culture Biotechnology |
| Zdroj: | Biotechnology and bioengineering. 106(1) |
| ISSN: | 1097-0290 |
| Popis: | A high rate of cell growth (micro) leading to rapid accumulation of viable biomass is a desirable phenotype during scale up operations and the early stages of production cultures. In order to identify genes and proteins that contribute to higher growth rates in Chinese hamster ovary (CHO) cells, a combined approach using microarray and proteomic expression profiling analysis was carried out on two matched pairs of CHO production cell lines that displayed either fast or slow growth rates. Statistical analysis of the microarray and proteomic data separately resulted in the identification of 118 gene transcripts and 58 proteins that were differentially expressed between the fast- and slow-growing cells. Overlap comparison of both datasets identified a priority list of 21 candidates associated with a high growth rate phenotype in CHO. Functional analysis (by siRNA) of five of these candidates identified the valosin-containing protein (VCP) as having a substantial impact on CHO cell growth and viability. Knockdown of HSPB1 and ENO1 also had an effect on cell growth (negative and positive, respectively). Further functional validation in CHO using both gene knockdown (siRNA) and overexpression (cDNA) confirmed that altered VCP expression impacted CHO cell proliferation, indicating that VCP and other genes and proteins identified here may play an important role in the regulation of CHO cell growth during log phase culture and are potential candidates for CHO cell line engineering strategies. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text