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Reverting cholesterol auxotrophy of NS0 cells by altering epigenetic gene silencing

2005; Wiley; Volume: 93; Issue: 4 Linguagem: Inglês

10.1002/bit.20720

1097-0290

Gargi Seth, Mustafa Öztürk, Wei‐Shou Hu,

RNA Research and Splicing

Biotechnology and BioengineeringVolume 93, Issue 4 p. 820-827 Communication to the Editor Reverting cholesterol auxotrophy of NS0 cells by altering epigenetic gene silencing Gargi Seth, Gargi Seth Department of Chemical Engineering and Materials Science, University of Minnesota, 421 Washington Avenue SE, Minneapolis, Minnesota; telephone: (612) 626-7630; fax: (612) 626-7246Search for more papers by this authorMustafa Ozturk, Mustafa Ozturk Department of Chemical Engineering and Materials Science, University of Minnesota, 421 Washington Avenue SE, Minneapolis, Minnesota; telephone: (612) 626-7630; fax: (612) 626-7246Search for more papers by this authorWei-Shou Hu, Corresponding Author Wei-Shou Hu [email protected] Department of Chemical Engineering and Materials Science, University of Minnesota, 421 Washington Avenue SE, Minneapolis, Minnesota; telephone: (612) 626-7630; fax: (612) 626-7246Department of Chemical Engineering and Materials Science, University of Minnesota, 421 Washington Avenue SE, Minneapolis, Minnesota; telephone: (612) 626-7630; fax: (612) 626-7246.Search for more papers by this author Gargi Seth, Gargi Seth Department of Chemical Engineering and Materials Science, University of Minnesota, 421 Washington Avenue SE, Minneapolis, Minnesota; telephone: (612) 626-7630; fax: (612) 626-7246Search for more papers by this authorMustafa Ozturk, Mustafa Ozturk Department of Chemical Engineering and Materials Science, University of Minnesota, 421 Washington Avenue SE, Minneapolis, Minnesota; telephone: (612) 626-7630; fax: (612) 626-7246Search for more papers by this authorWei-Shou Hu, Corresponding Author Wei-Shou Hu [email protected] Department of Chemical Engineering and Materials Science, University of Minnesota, 421 Washington Avenue SE, Minneapolis, Minnesota; telephone: (612) 626-7630; fax: (612) 626-7246Department of Chemical Engineering and Materials Science, University of Minnesota, 421 Washington Avenue SE, Minneapolis, Minnesota; telephone: (612) 626-7630; fax: (612) 626-7246.Search for more papers by this author First published: 27 September 2005 https://doi.org/10.1002/bit.20720Citations: 28AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Abstract NS0 is a cholesterol-requiring mouse myeloma cell line widely used in the production of recombinant antibodies. We have previously reported that the deficiency of 17β-hydroxysteroid dehydrogenase type7 (Hsd17b7) is responsible for the cholesterol auxotrophy of NS0 cells. Here we demonstrate DNA methylation to be the mechanism underlying transcriptional suppression of Hsd17b7 in cholesterol dependent NS0 cells. Analysis of the DNA methylation pattern revealed methylation of the CpG-rich region upstream of the Hsd17b7 transcription start site in NS0 cells. This is in contrast to the unmethylated status of this sequence in a naturally isolated cholesterol independent revertant cell population (NS0_r). This transcriptional repression was relieved after treating cells with the demethylating drug, 5-azacytidine. Drug treatment also gave rise to high frequency cholesterol-independent variants. Characterization of revertants revealed substantially elevated transcript level of 17β-hydroxysteroid dehydrogenase type7 (Hsd17b7) gene along with hypomethylation of the CpG-rich region. These results affirm that deficiency of Hsd17b7 causes cholesterol dependence of NS0 cells. Furthermore, induction of cholesterol independence by altering DNA methylation pattern alludes to the role of epigenetics in the metabolic adaptation of NS0 cells. With the widespread use of NS0 cells, this finding will have a significant impact on the optimization of recombinant antibody production processes. © 2005 Wiley Periodicals, Inc. References Antequera F. 2003. Structure, function, and evolution of CpG island promoters. Cell Mol Life Sci 60(8): 1647– 1658. Antequera F, Bird A. 1993. Number of CpG islands and genes in human and mouse. Proc Natl Acad Sci USA 90(24): 11995– 11999. Antequera F, Boyes J, Bird A. 1990. 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