Overexpression of GRP78 enhances survival of CHO cells in response to serum deprivation and oxidative stress
Overexpression of GRP78 enhances survival of CHO cells in response to serum deprivation and oxidative stress
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DOI：
10.1002/elsc.201500152
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发表时间：
2017-02-01
影响因子：
2.7
通讯作者：
Shen, Guanxin
中科院分区：
文献类型：
--
作者：
Jiang, Qing;Sun, Yuanli;Shen, Guanxin
中国仓鼠卵巢（CHO）细胞被认为是最常用的哺乳动物宿主之一，由于培养活力的丧失而降低了生产力。在 CHO 细胞中过度表达抗凋亡基因被开发为限制暴露于环境损伤时细胞死亡的一种手段。葡萄糖调节蛋白 78 (GRP78) 传统上被认为是参与蛋白质折叠和其他细胞过程的主要 ER 伴侣。它也是一种有效的抗凋亡蛋白，在细胞存活、增殖和转移中发挥着关键作用。在这项研究中，研究了 GRP78 对 CHO 细胞响应血清剥夺和氧化应激等环境损害的影响。首先，已证实 CHO 细胞对环境损害非常敏感。然后，建立过表达 GRP78 的 CHO 细胞系，并暴露于血清剥夺和 H2O2 中。结果表明，GRP78 工程增加了 CHO 细胞的活力并减少了细胞凋亡。 GRP78 工程带来的生存优势可以通过抑制参与应激细胞死亡途径的 caspase-3 来介导。此外，GRP78工程还提高了CHO细胞中抗转铁蛋白受体抗体的产量。 GRP78应该在生物制药行业有潜在的应用。
Chinese hamster ovary (CHO) cells are regarded as one of the most commonly used mammalian hosts, which decreases the productivity due to loss in culture viability. Overexpressing antiapoptosis genes in CHO cells was developed as a means of limiting cell death upon exposure to environmental insults. Glucose- regulated protein 78 (GRP78) is traditionally regarded as a major ER chaperone that participates in protein folding and other cell processes. It is also a potent antiapoptotic protein and plays a critical role in cell survival, proliferation, and metastasis. In this study, the impact of GRP78 on CHO cells in response to environmental insults such as serum deprivation and oxidative stress was investigated. First, it was confirmed that CHO cells were very sensitive to environmental insults. Then, GRP78 overexpressing CHO cell line was established and exposed to serum deprivation and H2O2. Results showed that GRP78 engineering increased the viability and decreased the apoptosis of CHO cells. The survival advantage due to GRP78 engineering could be mediated by suppression of caspase- 3 involved in cell death pathways in stressed cells. Besides, GRP78 engineering also enhanced yields of antibody against transferrin receptor in CHO cells. GRP78 should be a potential application in the biopharmaceutical industries.