A DIGE Approach for the Assessment of Differential Expression of the CHO Proteome Under Sodium Butyrate Addition: Effect of Bcl-x(L) Overexpression

Abstract

Bcl-x(L), a member of the Bcl-2 family, is known to inhibit apoptosis of recombinant Chinese hamster ovary (rCHO) cells induced by the addition of sodium butyrate (NaBu), which is used for the elevated expression of recombinant protein. In order to understand the intracellular effects of Bcl-x(L), overexpression on CHO cells treated with NaBu, changes to the proteome caused by controlled Bcl-x(L) expression in rCHO cells producing erythropoietin (EPO) in,the presence of 3 mM NaBu were evaluated using two-dimensional differential in-gel electrophoresis (2D-DIGE) and MS analysis. The consequences of Bcl-x(L) overexpression were not limited to the apoptotic signaling pathway. Out of eight proteins regulated significantly by Bcl-x(L) overexpression in 3 mM NaBu addition culture, four proteins were related to cell survival (Iq motif-containing GTPase-activating protein 1), cell proliferation (dihydrolipoamide-S-acetyltransferase, guanine nucleotide binding protein alpha interacting 2), and repair of DNA damage (BRCA and CDKN1A interacting protein). Taken together, a DIGE approach reveals that overexpression of Bcl-x(L) not only inhibits apoptosis in the presence of NaBu but also affects cell proliferation and survival in various aspects. Biotechnol. Bioeng. 2010;105: 358-367. (C) 2009 Wiley Periodicals, Inc.