Transcriptome and proteome analysis of Chinese hamster ovary cells under low temperature and butyrate treatment
Transcriptome and proteome analysis of Chinese hamster ovary cells under low temperature and butyrate treatment
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DOI：
10.1016/j.jbiotec.2009.09.008
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发表时间：
2010-01-15
影响因子：
4.1
通讯作者：
Hu, Wei-Shou
中科院分区：
文献类型：
--
作者：
Kantardjieff, Anne;Jacob, Nitya M.;Hu, Wei-Shou
选择高生产力的重组中国仓鼠卵巢 (CHO) 细胞能够分泌免疫球蛋白 G (IgG) 分子，其水平可与体内浆细胞相媲美。在 33 摄氏度下进行丁酸盐处理后，观察到生产率进一步提高。为了更好地了解导致生产率提高的机制，使用寡 DNA 微阵列研究了经过这些处理的抗体产生细胞系的转录反应。通过距离计算，超过 900 个基因被鉴定为在丁酸盐处理的 33°C 培养物和未处理的培养物之间存在动力学差异表达。此外，治疗后重链和轻链 IgG 基因的转录水平增加。使用稳定同位素标记 (SILAC)，通过跟踪切换至未标记介质后同位素标记的衰减来研究 IgG 的分泌率。处理和未处理的培养物都表现出非常相似的 IgG 分泌动力学。相反，治疗后发现细胞内 IgG 含量升高。这一结果表明，治疗下生产力的提高可归因于细胞分泌能力的提高，而不是分泌途径中停留时间的缩短。基因集富集分析 (GSEA) 的结果进一步支持了这一假设，该分析揭示了分泌途径的元素，包括高尔基体、细胞骨架蛋白结合和小 GTP 酶介导的信号转导，都得到富集，因此可能在 33 摄氏度丁酸盐处理下观察到的重组蛋白产量增加中发挥作用。(C) 2009 Elsevier B.V. 保留所有权利。
Recombinant Chinese hamster ovary (CHO) cells selected for high productivity are capable of secreting immunoglobulin G (IgG) molecules at a level that rivals plasma cells in vivo. Following butyrate treatment at 33 degrees C, further increases in productivity are observed. To better understand the mechanisms by which this increased productivity is incurred, the transcriptional response of an anti body-producing cell line undergoing these treatments was investigated using oligo-DNA microarrays. Using distance calculations, more than 900 genes were identified as kinetically differentially expressed between the butyrate-treated 33 degrees C culture and the untreated culture. Furthermore, transcript levels of the heavy and light chain IgG genes increased following treatment. Using stable isotope labeling (SILAC), the secretion rate of IgG was investigated by tracking the decay of the isotope label upon switching to unlabeled medium. Both treated and untreated cultures exhibited very similar IgG secretion kinetics. In contrast, the intracellular IgG content was found to be elevated following treatment. This result suggests that increased productivity under treatment is attributable to elevated cellular secretory capacity, rather than shorter holding times in the secretory pathway. This hypothesis is further supported by the results of gene set enrichment analysis (GSEA), which revealed that elements of the secretory pathway, including Golgi apparatus, cytoskeleton protein binding and small GTPase-mediated signal transduction are enriched and thus may play a role in the increased recombinant protein production observed under butyrate treatment at 33 degrees C. (C) 2009 Elsevier B.V. All rights reserved.