The effect of oxidative stress on the production of the recombinant protein, interferon γ, produced by Chinese hamster ovary cells in stirred-batch culture

1997 ◽  
Vol 48 (2) ◽  
pp. 198-203 ◽  
Author(s):  
C. A. Dunster ◽  
K. H. Cheeseman ◽  
S. P. Maddix
Keyword(s):  
Oxidative Stress ◽  
Recombinant Protein ◽  
Batch Culture ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Interferon Γ ◽  
Ovary Cells

scholarly journals Increased sensitivity to oxidative injury in chinese hamster ovary cells stably transfected with rat liver S-adenosylmethionine synthetase cDNA

1996 ◽  
Vol 319 (3) ◽  
pp. 767-773 ◽  
Author(s):  
Estrella SÁNCHEZ-GÓNGORA ◽  
John G. PASTORINO ◽  
Luis ALVAREZ ◽  
María A. PAJARES ◽  
Concepción GARCÍA ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Rat Liver ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Wild Type ◽  
Ovary Cells ◽  
Transfected Cells ◽  
In The Wild ◽  
Adenosylmethionine Synthetase

Chinese hamster ovary cells were stably transfected with rat liver S-adenosylmethionine synthetase cDNA. As a result, S-adenosylmethionine synthetase activity increased 2.3-fold, an effect that was accompanied by increased S-adenosylmethionine, a depletion of ATP and NAD levels, elevation of the S-adenosylmethionine/S-adenosylhomocysteine ratio (the methylation ratio), increased DNA methylation and polyamine levels (spermidine and spermine), and normal GSH levels. By contrast, the transfected cells showed normal growth curves and morphology. Exposure to an oxidative stress by the addition of H2O2 resulted in a greater consumption of ATP and NAD in the transfected cells than in the wild-type cells. In turn, cell killing by H2O2 was greater in the transfected cells than in the wild-type cells. This killing of Chinese hamster ovary cells by H2O2 involved the activation of poly(ADP-ribose) polymerase with the resultant loss of NAD and ATP. 3-Aminobenzamide, an inhibitor of poly(ADP-ribose) polymerase, but not the antioxidant N,N´-diphenylphenylenediamine, prevented the killing of Chinese hamster ovary cells by H2O2 and maintained the contents of NAD and ATP. The results of this study indicate that a moderate activation of the synthesis of S-adenosylmethionine leads to ATP and NAD depletion and to a greater sensitivity to cell killing by oxidative stress.

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