Gadd45-induced cell cycle G2/M arrest for improved transient gene expression in Chinese hamster ovary cells
Gadd45-induced cell cycle G2/M arrest for improved transient gene expression in Chinese hamster ovary cells
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Gadd45 诱导细胞周期 G2/M 停滞以改善中国仓鼠卵巢细胞的瞬时基因表达
DOI：
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发表时间：
2014
影响因子：
3.2
通讯作者：
G. Lee
中科院分区：
文献类型：
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作者：
Won Hee Kim;Yeon Jung Kim;G. Lee
Gadd45 是一种 p53 调节蛋白，参与细胞周期 G2/M 期停滞。为了改善中国仓鼠卵巢 (CHO) 细胞中的瞬时基因表达 (TGE)，我们使用编码 Fcfusion 蛋白和重组抗体的两种不同表达载体研究了 Gadd45 诱导的细胞周期停滞对 CHO 细胞中 TGE 的影响。为了调节CHO细胞中Gadd45的表达，使用多西环素控制的T-REx系统建立了CHO-TREx-gadd45细胞系。在 TGE 培养过程中，Gadd45 过表达严重抑制细胞生长，但显着增强 TGE。与未过表达Gadd45的培养物相比，Fc融合蛋白和人源化抗体的TGE分别增加了111%和93%。尽管 Gadd45 过表达导致细胞生长停滞，但 TGE 的增强是由于转染效率提高、细胞大小增加和主动 DNA 去甲基化导致比生产率显着提高。综上所述，此处获得的数据表明，Gadd45 诱导的 G2/M 期细胞周期停滞可以显着增强 CHO 细胞中的 TGE。
Gadd45 is a p53-regulated protein and is involved in cell cycle arrest in the G2/M phase. In an effort to improve transient gene expression (TGE) in Chinese hamster ovary (CHO) cells, the effect of Gadd45-induced cell cycle arrest on TGE in CHO cells was investigated using the two different expression vectors encoding Fcfusion protein and recombinant antibody. To regulate the expression of Gadd45 in CHO cells, the CHO-TREx-gadd45 cell line was established using the T-REx system controlled by doxycycline. During the cultures for TGE, Gadd45 overexpression severely inhibited cell growth, but significantly enhanced TGE. Compared with the culture without Gadd45 overexpression, the TGE of Fc fusion protein and humanized antibody were increased by 111 and 93%, respectively. The enhanced TGE, despite the cell growth arrest induced by Gadd45 overexpression, was due to the significantly increased specific productivity, resulting from enhanced transfection efficiency, increased cell size, and active DNA demethylation. Taken together, the data obtained here demonstrate that Gadd45-induced cell cycle arrest in G2/M phase can significantly enhance TGE in CHO cells.