title: A mechanistic study on the effect of dexamethasone in moderating cell death in Chinese Hamster Ovary cell cultures.
abstract: Dexamethasone (DEX) was previously shown (Jing et al., Biotechnol Bioeng. 2010;107:488-496) to play a dual role in increasing sialylation of recombinant glycoproteins produced by Chinese Hamster Ovary (CHO) cells. DEX addition increased sialic acid levels of a recombinant fusion protein through increased expression of α2,3-sialyltransferase and β1,4-galactosyltransferase, but also decreased the sialidase-mediated, extracellular degradation of sialic acid through slowing cell death at the end of the culture period. This study examines the underlying mechanism for this cytoprotective action by studying the transcriptional response of the CHO cell genome upon DEX treatment using DNA microarrays and gene ontology term analysis. Many of those genes showing a significant transcriptional response were associated with the regulation of programmed cell death. The gene with the highest change in expression level, as validated by Quantitative PCR assays with TaqMan® probes and confirmed by Western Blot analysis, was the antiapoptotic gene Tsc22d3, also referred to as GILZ (glucocorticoid-induced leucine zipper). The pathway by which DEX suppressed cell death towards the end of the culture period was also confirmed by showing involvement of glucocorticoid receptors and GILZ through studies using the glucocorticoid antagonist mifepristone (RU-486). These findings advance the understanding of the mechanism by which DEX suppresses cell death in CHO cells and provide a rationale for the application of glucocorticoids in CHO cell culture processes.
authors.affiliation: Biologics Process and Product Development, Technical Operations, Bristol-Myers Squibb Company, East Syracuse, NY 13057, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Jing
authors.forename: Yueming
authors.initials: Y
authors.lastname: Qian
authors.forename: Mahmoud
authors.initials: M
authors.lastname: Ghandi
authors.forename: Aiqing
authors.initials: A
authors.lastname: He
authors.forename: Michael C
authors.initials: MC
authors.lastname: Borys
authors.forename: Shih-Hsie
authors.initials: SH
authors.lastname: Pan
authors.forename: Zheng Jian
authors.initials: ZJ
authors.lastname: Li
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D016466:CHO Cells; D018929:Cell Culture Techniques; D016923:Cell Death / Q000187:drug effects; D002477:Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D006224:Cricetinae; D003907:Dexamethasone / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D051379:Mice; D011965:Receptors, Glucocorticoid / Q000235:genetics / Q000378:metabolism
chemical_list: D011965:Receptors, Glucocorticoid; D003907:Dexamethasone