Calnexin overexpression sensitizes recombinant CHO cells to apoptosis induced by sodium butyrate treatment

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Sodium butyrate (NaBu) can enhance the expression of foreign genes in recombinant Chinese hamster ovary (rCHO) cells, but it can also inhibit cell growth and induce cellular apoptosis. In this study, the potential role of calnexin (Cnx) expression in rCHO cells treated with 5 mM NaBu was investigated for rCHO cells producing tumor necrosis factor receptor FC. To regulate the Cnx expression level, a tetracycline-inducible system was used. Clones with different Cnx expression levels were selected and investigated. With regard to productivity per cell (q (p)), NaBu enhanced the q (p) by over twofold. Under NaBu treatment, Cnx overexpression further enhanced the q (p) by about 1.7-fold. However, under NaBu stress, the cells overexpressing Cnx showed a poorer viability profile with a consistent difference of over 25% in the viability when compared to the Cnx-repressed condition. This drop in the viability was attributed to increased apoptosis seen in these cells as evidenced by enhanced poly (ADP-ribose) polymerase cleavage and cytochrome C release. Ca2+ localization staining and subsequent confocal imaging revealed elevated cytosolic Ca2+ ([Ca2+](c)) in the Cnx-overexpressing cells when compared to the Cnx-repressed condition, thus endorsing the increased apoptosis observed in these cells. Taken together, Cnx overexpression not only improved the q (p) of cells treated with NaBu, but it also sensitized cells to apoptosis.
Publisher
SPRINGER
Issue Date
2009-01
Language
English
Article Type
Article
Keywords

HAMSTER OVARY CELLS; STRESS-INDUCED APOPTOSIS; HIGH-LEVEL EXPRESSION; ENDOPLASMIC-RETICULUM; MOLECULAR CHAPERONE; GENE-EXPRESSION; THROMBOPOIETIN PRODUCTIVITY; SURFACE EXPRESSION; MAMMALIAN-CELLS; BATCH CULTURES

Citation

CELL STRESS & CHAPERONES, v.14, no.1, pp.49 - 60

ISSN
1355-8145
DOI
10.1007/s12192-008-0054-0
URI
http://hdl.handle.net/10203/100878
Appears in Collection
BS-Journal Papers(저널논문)
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