De Leon Gatti Marcela, Wlaschin Katie F, Nissom Peter Morin, Yap Miranda, Hu Wei-Shou
Department of Chemical Engineering and Materials Science, University of Minnesota, 421 Washington Ave SE., Minneapolis, MN 55455-0132, USA.
J Biosci Bioeng. 2007 Jan;103(1):82-91. doi: 10.1263/jbb.103.82.
DNA microarray based transcriptome analysis has become widely used in biomedical research; however, the lack of DNA sequence information available for Chinese hamster ovary (CHO) cells has hampered the application of microarrays for this cell line widely used for recombinant therapeutic protein production. We have constructed an expressed sequence tag (EST) based CHO DNA microarray and employed it for comparative transcriptome analysis of CHO cells and mouse hybridoma cells treated with sodium butyrate. Cross-species hybridization of CHO transcripts to mouse DNA microarrays was also performed to assess the utility of cross-species microarray. The average identity among probe sequences present on both the CHO and mouse microarray was 89.6%. Although cross-species hybridization yielded non-contradicting results when compared with the same-species arrays, decreased sensitivity was observed and resulted in fewer differentially expressed genes being confidently identified. The comparatively small number of genes probed using the CHO microarray and the low number of genes identified as differentially expressed in the cross-species hybridization limited physiological interpretation of the response of CHO cells to sodium butyrate treatment. Nevertheless, when all results are combined, mouse hybridoma and CHO cells can be seen to respond similarly to butyrate treatment, affecting histone modification, chaperones, lipid metabolism, and protein processing. To further develop the utility of microarray technology in cell culture process development, an expansion of current CHO cell sequencing efforts to increase the coverage of genes on available microarrays is warranted.
基于DNA微阵列的转录组分析已在生物医学研究中广泛应用;然而,由于缺乏中国仓鼠卵巢(CHO)细胞的DNA序列信息,阻碍了微阵列在这种广泛用于重组治疗性蛋白质生产的细胞系中的应用。我们构建了基于表达序列标签(EST)的CHO DNA微阵列,并将其用于丁酸钠处理的CHO细胞和小鼠杂交瘤细胞的比较转录组分析。还进行了CHO转录本与小鼠DNA微阵列的跨物种杂交,以评估跨物种微阵列的实用性。CHO和小鼠微阵列上存在的探针序列之间的平均一致性为89.6%。虽然与同物种阵列相比,跨物种杂交产生的结果并不矛盾,但观察到灵敏度降低,导致可靠鉴定的差异表达基因数量减少。使用CHO微阵列检测的基因数量相对较少,以及在跨物种杂交中鉴定为差异表达的基因数量较少,限制了对CHO细胞对丁酸钠处理反应的生理学解释。尽管如此,当所有结果综合起来时,可以看出小鼠杂交瘤细胞和CHO细胞对丁酸盐处理的反应相似,影响组蛋白修饰、伴侣蛋白、脂质代谢和蛋白质加工。为了进一步开发微阵列技术在细胞培养工艺开发中的实用性,有必要扩大当前CHO细胞测序工作,以增加现有微阵列上基因的覆盖范围。