Gene transcription acceleration: main cause of hepatitis B surface antigen production improvement by dimethyl sulfoxide in the culture of Chinese hamster ovary cells

Abstract

The production and specific productivity of hepatitis B surface antigen (HBsAg) in recombinant Chinese hamster ovary (CHO) cells were increased by 81% and threefold, respectively, when supplemented with 1.5% dimethyl sulfoxide (DMSO) in the culture medium. To investigate the mechanism of DMSO effect on HBsAg production improvement, HBsAg mRNA level was measured by real-time PCR. HBsAg mRNA was increased by about 1.5-fold at 1.5% DMSO. The increase could derive from the increase of HBsAg gene copy number, the improvement of HBsAg mRNA stability, or the acceleration of HBsAg gene transcription. It was found that HBsAg gene copy number was not significantly changed in the cells stimulated with DMSO. HBsAg mRNA stability of cells with DMSO treatment was also not obviously different from control, and the mRNA half-life of 5.58 h in the cells at 1.5% DMSO was comparable to that of 5.36 h in the control culture. DMSO resulted in 80% increase in HBsAg gene transcription activity assessed using a nuclear run-on transcription assay. It could be deduced that the acceleration of HBsAg gene transcription is the main cause of HBsAg production improvement.

Production and characterization of active recombinant interleukin‐12/eGFP fusion protein in stably‐transfected DF1 chicken cells

“…10,11 Thus, these results and the present study on the rchIL-12/eGFP protein suggests that specific gene expression patterns after transfection may depend on the host cell targets. In a study on the effect of DMSO on transcription level of HBsAg gene expression, Wang et al (2006) 19 indicated that some post-translation events, such as the saturation of the secretion pathway, the inhibition of DMSO on HBsAg secretion pathway, and the limitation of post-translation modification of protein, could involve in restriction on the HBsAg secretion. For example, DMSO downregulates HSP70 and ERP57, which play an important role in stabilizing the folding and assembly intermediates of newly synthesized polypeptides.…”

“…12,15,16 DMSO may have a broad impact on many aspects of gene expression in eukaryotic cell systems. 15,16,19,21 However, little is known about the exact mechanism by which DMSO exerts an effect. It has been demonstrated that the production of hepatitis B surface antigen (HBsAg) in recombinant CHO cells was increased by 81% and the productivity of HBsAg by a single cell was increased by threefold when the culture medium was supplemented with 1.5% DMSO.…”

“…The main cause for the increase in HBsAg production might be the acceleration of HBsAg gene transcription. 19 Recently, a study reported that DMSO can increase the expression of recombinant cyclooxyenase 1 (COX-1) in stably transfected insect S2 cells. 22 DMSO increased the expression of nitric oxide synthase (NOS), which is closely related with the synthesis of recombinant COX-1 mRNA.…”

Enhanced production of recombinant proteins by a small molecule protein synthesis enhancer in combination with an antioxidant in recombinant Chinese hamster ovary cells

“…Bioactive chemicals can target many cellular mechanisms, with inducing upregulation of protein production. A wide variety of molecules have been used to accomplish this, including quinoline thioethers [12], dimethyl sulfoxide [3,7,[13][14][15], and aurintricarboxylic acid [7]. Additionally, broad classes of small molecules including aromatic carboxylic acids, hydroxamic acids, and acetamides have been studied [16].…”

Enhanced expression of recombinant human cyclooxygenase 1 from stably-transfected Drosophila melanogaster S2 cells by dimethyl sulfoxide is mediated by up-regulation of nitric oxide synthase and transcription factor Kr-h1

“…DMSO induces apoptosis in some cell lines (Marthyn et al 1998) whereas it inhibits cell density-dependent apoptosis of CHO cells (Fiore and Degrassi 1999). DMSO may have a broad impact on many aspects of gene expression in animal cells (Li et al 2006;Liu et al 2001;Wang et al 2007;Sumida et al 2011). DMSO can stimulate CHO cells to synthesize different recombinant proteins .…”